Conformationally restricted urea inhibitors of soluble epoxide hydrolase

ABSTRACT

Inhibitors of the soluble epoxide hydrolase (sEH) are provided that incorporate multiple pharmacophores and are useful in the treatment of diseases.

CROSS-REFERENCES TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional PatentApplication No. 60/782,172, filed Mar. 13, 2006, which is incorporatedby reference herein in its entirety.

STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSOREDRESEARCH OR DEVELOPMENT

The U.S. Government has certain rights to the invention pursuant tocontract ES02710 & HL078096 awarded by the National Institutes ofHealth.

BACKGROUND OF THE INVENTION

Epoxide hydrolases (EHs, EC 3.3.2.3) catalyze the hydrolysis of epoxidesor arene oxides to their corresponding diols by the addition of water(see, Oesch, F., et al., Xenobiotica 1973, 3, 305-340). Some EHs play animportant role in the metabolism of a variety of compounds includinghormones, chemotherapeutic drugs, carcinogens, environmental pollutants,mycotoxins, and other harmful foreign compounds.

There are two well-studied EHs, microsomal epoxide hydrolase (mEH) andsoluble epoxide hydrolase (sEH). These enzymes are very distantlyrelated, have different subcellular localization, and have different butpartially overlapping substrate selectivities. The soluble andmicrosomal EH forms are known to complement each other in degrading someplant natural products (see, Hammock, B. D., et al., COMPREHENSIVETOXICOLOGY. Oxford: Pergamon Press 1977, 283-305 and Fretland, A. J., etal., Chem. Biol. Intereract 2000, 129, 41-59).

The major role of the sEH is in the metabolism of lipid epoxidesincluding the metabolism of arachidonic acid (see, Zeldin, D. C., etal., J. Biol. Chem. 1993, 268, 6402-6407), linoleic acid (see,Moghaddam, M. F., et al., Nat. Med. 1997, 3, 562-567) acid, some ofwhich are endogenous chemical mediators (see, Carroll, M. A., et al.,Thorax 2000, 55, S13-16). Epoxides of arachidonic acid(epoxyeicosatrienoic acids or EETs) and other lipid epoxides and diolsare known effectors of blood pressure (see, Capdevila, J. H., et al., J.Lipid. Res. 2000, 41, 163-181), and modulators of vascular permeability(see, Oltman, C. L., et al., Circ Res. 1998, 83, 932-939). Thevasodilatory properties of EETs are associated with an increasedopen-state probability of calcium-activated potassium channels leadingto hyperpolarization of the vascular smooth muscle (see Fisslthaler, B.,et al., Nature 1999, 401, 493-497). Hydrolysis of the arachidonateepoxides by sEH diminishes this activity (see, Capdevila, J. H., et al.,J. Lipid. Res. 2000, 41, 163-181). sEH hydrolysis of EETs also regulatestheir incorporation into coronary endothelial phospholipids, suggestinga regulation of endothelial function by sEH (see, Weintraub, N. L., etal., Am. J. Physiol. 1992, 277, H2098-2108). It has recently been shownthat treatment of spontaneous hypertensive rats (SHRs) with selectivesEH inhibitors significantly reduces their blood pressure (see, Yu, Z.,et al., Circ. Res. 2000, 87, 992-998). In addition, it was claimed thatmale knockout sEH mice have significantly lower blood pressure thanwild-type mice (see Sinal, C. J., et al., J. Biol. Chem. 2000, 275,40504-405010), however subsequent studies demonstrated with backbreeding into C57b mice that 20-HETE levels increased compensating forthe increase in plasma EETs (see, Luria, A. et al., J. Biol. Chem. 2007,282:2891-2898.

The EETs have also demonstrated anti-inflammatory properties inendothelial cells (see, Node, K., et al., Science 1999, 285, 1276-1279and Campbell, W. B. Trends Pharmacol. Sci. 2000, 21, 125-127). Incontrast, diols derived from epoxy-linoleate (leukotoxin) perturbmembrane permeability and calcium homeostasis (see, Moghaddam, M. F., etal., Nat. Med. 1997, 3, 562-567), which results in inflammation that ismodulated by nitric oxide synthase and endothelin-1 (see, Ishizaki, T.,et al., Am. J. Physiol. 1995, 269, L65-70 and Ishizaki, T., et al., J.Appl. Physiol. 1995, 79, 1106-1611). Micromolar concentrations ofleukotoxin reported in association with inflammation and hypoxia (see,Dudda, A., et al., Chem. Phys. Lipids 1996, 82, 39-51), depressmitochondrial respiration in vitro (see, Sakai, T., et al., Am. J.Physiol. 1995, 269, L326-331), and cause mammalian cardiopulmonarytoxicity in vivo (see, Ishizaki, T., et al., Am. J. Physiol. 1995, 269,L65-70; Fukushima, A., et al., Cardiovasc. Res. 1988, 22, 213-218; andIshizaki, T., et al., Am. J. Physiol. 1995, 268, L123-128). Leukotoxintoxicity presents symptoms suggestive of multiple organ failure andacute respiratory distress syndrome (ARDS) (see, Ozawa, T. et al., Am.Rev. Respir. Dis. 1988, 137, 535-540). In both cellular and organismalmodels, leukotoxin-mediated toxicity is dependent upon epoxidehydrolysis (see, Moghaddam, M. F., et al., Nat. Med. 1997, 3, 562-567;Morisseau, C., et al., Proc. Natl. Acad. Sci. USA 1999, 96, 8849-8854;and Zheng, J., et al., Am. J. Respir. Cell Mol. Biol. 2001, 25,434-438), suggesting a role for sEH in the regulation of inflammationand vascular permeability. The bioactivity of these epoxy-fatty acidssuggests that inhibition of vicinal-dihydroxy-lipid biosynthesis mayhave therapeutic value, making sEH a promising pharmacological target.

Recently, 1,3-disubstituted ureas, carbamates, and amides have beenreported as new potent and stable inhibitors of sEH See, U.S. Pat. No.6,150,415. Compounds 192 and 686 are representative structures for thistype of inhibitors (FIG. 1, therein). These compounds are competitivetight-binding inhibitors with nanomolar K_(I) values that interactstoichiometrically with purified recombinant sEH (see, Morisseau, C., etal., Proc. Natl. Acad. Sci. USA 1999, 96, 8849-8854). Based on the X-raycrystal structure, the urea inhibitors were shown to establish hydrogenbonds and to form salt bridges between the urea function of theinhibitor and residues of the sEH active site, mimicking featuresencountered in the reaction coordinate of epoxide ring opening by thisenzyme (see, Argiriadi, M. A., et al., Proc. Natl. Acad. Sci. USA 1999,96, 10637-10642 and Argiriadi, M. A., et al., J. Biol. Chem. 2000, 275,15265-15270). These inhibitors efficiently reduced epoxide hydrolysis inseveral in vitro and in vivo models (see, Yu, Z., et al., Circ. Res.2000, 87, 992-998; Morisseau, C., et al., Proc. Natl. Acad. Sci. USA1999, 96, 8849-8854; and Newman, J. W., et al., Environ. HealthPerspect. 2001, 109, 61-66). Despite the high activity associated withthese inhibitors, there exists a need for compounds possessing similaror increased activities, preferably with improved solubility and/orpharmacokinetic properties to facilitate formulation and delivery.

The present invention provides such compounds along with methods fortheir use and compositions that contain them.

BRIEF SUMMARY OF THE INVENTION

In one aspect, the present invention provides a method for inhibiting asoluble epoxide hydrolase, comprising contacting the soluble epoxidehydrolase with an inhibiting amount of a compound having the formula(I):

The symbol R¹ is a member selected from the group consisting ofC₁-C₈alkyl, arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each ofwhich is optionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl, andheteroaryl; wherein said cyclic portions are monocyclic or polycyclic.In one embodiment, the 1 to 2 substituents are each independentlyselected from the group consisting of C₁-C₈alkyl and C₁-C₈alkoxy. In oneembodiment, the 1 to 2 substituents are each independently selected fromthe group consisting of C₁-C₈haloalkyl and C₁-C₈haloalkoxy.

The symbol Y¹ is selected from the group consisting of a bond, C(R⁵)₂,NR⁵ and O.

The symbol Y² is selected from the group consisting of a bond, NR⁵ andO.

Each symbol R², R³ and R⁵ is independently selected from the groupconsisting of H, C₁-C₈alkyl and COR⁶.

The symbol A is heterocyclyl, optionally substituted with from 1 to 2 R⁷substituents.

The symbol L is selected from the group consisting of a direct bond,C₁-C₁₂alkylene, C₁-C₁₂heteroalkylene, C₃-C₆cycloalkylene, arylene,heteroarylene, —CO—, —SO_(m)— and —Se—.

The symbol R⁴ is selected from the group consisting of H, C₁-C₈alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₈heteroalkyl, arylC₀-C₈alkyl,C₃-C₁₂cycloalkyl and heterocyclyl, each of which is optionallysubstituted. In one embodiment, each C₁-C₈alkyl, C₂-C₆alkenyl,C₂-C₆alkynyl, C₁-C₈heteroalkyl, arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl andheterocyclyl group is optionally substituted with from 1 to 2substituents each independently selected from the group consisting ofC₁-C₈alkyl, halo, C₁-C₈heteroalkyl, arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ andheteroaryl. In one embodiment, R⁴ is selected from the group consistingof C₁-C₈alkyl and C₁-C₈alkoxy. In one embodiment, R⁴ is selected fromthe group consisting of C₁-C₈haloalkyl and C₁-C₈haloalkoxy.

Each symbol R⁶ is independently selected from the group consisting of H,C₁-C₈alkyl, OH, C₁-C₈alkoxy and amino.

Each symbol R⁷ is selected from the group consisting of halo, nitro,C₁-C₈alkyl, C₁-C₈alkylamino, hydroxyC₁-C₈alkyl, haloC₁-C₈alkyl,carboxyl, hydroxyl, C₁-C₈alkoxy, C₁-C₈alkoxyC₁-C₈alkoxy,haloC₁-C₈alkoxy, thioC₁-C₈alkyl, aryl, aryloxy, C₃-C₈cycloalkyl,C₃-C₈cycloalkyl C₁-C₈alkyl, heteroaryl, arylC₁-C₈alkyl,heteroarylC₁-C₈alkyl, C₂-C₈alkenyl containing 1 to 2 double bonds,C₂-C₈alkynyl containing 1 to 2 triple bonds, C₄-C₈alk(en)(yn)yl groups,cyano, formyl, C₁-C₈alkylcarbonyl, arylcarbonyl, heteroarylcarbonyl,C₁-C₈alkoxycarbonyl, aryloxycarbonyl, aminocarbonyl,C₁-C₈alkylaminocarbonyl, C₁-C₈dialkylaminocarbonyl, arylaminocarbonyl,diarylaminocarbonyl, arylC₁-C₈alkylaminocarbonyl, haloC₁-C₈alkoxy,C₂-C₈alkenyloxy, C₂-C₈alkynyloxy, arylC₁-C₈alkoxy, aminoC₁-C₈alkyl,C₁-C₈alkylaminoC₁-C₈alkyl, C₁-C₈dialkylaminoC₁-C₈alkyl,arylaminoC₁-C₈alkyl, amino, C₁-C₈dialkylamino, arylamino,arylC₁-C₈alkylamino, C₁-C₈alkylcarbonylamino, arylcarbonylamino, azido,mercapto, C₁-C₈alkylthio, arylthio, haloC₁-C₈alkylthio, thiocyano,isothiocyano, C₁-C₈alkylsulfinyl, C₁-C₈alkylsulfonyl, arylsulfinyl,arylsulfonyl, aminosulfonyl, C₁-C₈alkylaminosulfonyl,C₁-C₈dialkylaminosulfonyl and arylaminosulfonyl.

The subscript n is an integer of 0 to 1.

The subscript m is an integer of from 0 to 2.

The compounds include all pharmaceutically acceptable derivativesthereof, such as salts, prodrugs, soft drugs, solvates and hydrates.

In a related aspect, the present invention provides methods of treatingdiseases modulated by soluble epoxide hydrolases, the method comprisingadministering to a subject in need of such treatment an effective amountof a compound having a formula selected from formula (I), above. In oneaspect, the effective amount is a therapeutically effective amount.

In other aspects, the present invention provides methods of reducingrenal deterioration in a subject, the method comprising administering tothe subject an effective amount of a compound of formula (I), above.

In a related aspect, the present invention provides methods method forinhibiting progression of nephropathy in a subject, the methodcomprising administering to the subject an effective amount of acompound of formula (I), above.

In another aspect, the present invention provides for reducing bloodpressure in a subject, the method comprising administering to thesubject an effective amount of a compound of formula (I), above.

In a related aspect, the present invention provides methods ofinhibiting the proliferation of vascular smooth muscle cells in asubject, the method comprising administering to the subject an effectiveamount of a compound of formula (I), above.

In another aspect, the present invention provides methods of inhibitingthe progression of an obstructive pulmonary disease, an interstitiallung disease, or asthma in a subject, the method comprisingadministering to the subject an effective amount of a compound offormula (I), above. The obstructive pulmonary disease can be, forexample, chronic obstructive pulmonary disease (“COPD”), emphysema, orchronic bronchitis. The interstitial lung disease can be, for example,idiopathic pulmonary fibrosis, or one associated with occupationalexposure to a dust.

In yet another aspect, the present invention provides compounds having aformula (I) above, as well as pharmaceutical compositions containing oneor more of the subject compounds.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. Pharmacokinetic profile of compounds with piperidinesubstitutions given as a single oral dose of 0.3 mg/kg.

FIG. 2. Pharmacokinetic profile of compounds 1153, 1155 and 1645.Compounds were administered orally to canines at 0.3 mg/kg.

FIG. 3. Pharmacokinetic profile of compound 1153 following single oraladministration 0.1 and 0.3 mg/kg orally.

FIG. 4. Pharmacokinetic profile of compound 1153 and other compoundsfollowing single oral administration 0.3 mg/kg of canine model.

FIG. 5. Exposure of selected compounds as a function of inverse potency.

DETAILED DESCRIPTION OF THE INVENTION

Abbreviations and Definitions:

“cis-Epoxyeicosatrienoic acids” (“EETs”) are biomediators synthesized bycytochrome P450 epoxygenases.

“Epoxide hydrolases” (“EH;” EC 3.3.2.3) are enzymes in the alpha/betahydrolase fold family that add water to 3 membered cyclic ethers termedepoxides.

“Soluble epoxide hydrolase” (“sEH”) is an enzyme which in endothelial,smooth muscle and other cell types converts EETs to dihydroxyderivatives called dihydroxyeicosatrienoic acids (“DHETs”). The cloningand sequence of the murine sEH is set forth in Grant et al., J. Biol.Chem. 268(23):17628-17633 (1993). The cloning, sequence, and accessionnumbers of the human sEH sequence are set forth in Beetham et al., Arch.Biochem. Biophys. 305(1):197-201 (1993). The amino acid sequence ofhuman sEH is also set forth as SEQ ID NO:2 of U.S. Pat. No. 5,445,956;the nucleic acid sequence encoding the human sEH is set forth asnucleotides 42-1703 of SEQ ID NO:1 of that patent. The evolution andnomenclature of the gene is discussed in Beetham et al., DNA Cell Biol.14(1):61-71 (1995). Soluble epoxide hydrolase represents a single highlyconserved gene product with over 90% homology between rodent and human(Arand et al., FEBS Lett., 338:251-256 (1994)).

The terms “treat”, “treating” and “treatment” refer to any method ofalleviating or abrogating a disease or its attendant symptoms.

The term “therapeutically effective amount” refers to that amount of thecompound being administered sufficient to prevent or decrease thedevelopment of one or more of the symptoms of the disease, condition ordisorder being treated.

The term “modulate” refers to the ability of a compound to increase ordecrease the function, or activity, of the associated activity (e.g.,soluble epoxide hydrolase). “Modulation”, as used herein in its variousforms, is meant to include antagonism and partial antagonism of theactivity associated with sEH. Inhibitors of sEH are compounds that,e.g., bind to, partially or totally block the enzyme's activity.

The term “compound” as used herein is intended to encompass not only thespecified molecular entity but also its pharmaceutically acceptable,pharmacologically active derivatives, including, but not limited to,salts, prodrug conjugates such as esters and amides, metabolites,hydrates, solvates and the like.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients in the specified amounts, as wellas any product which results, directly or indirectly, from combinationof the specified ingredients in the specified amounts. By“pharmaceutically acceptable” it is meant the carrier, diluent orexcipient must be compatible with the other ingredients of theformulation and not deleterious to the recipient thereof.

The “subject” is defined herein to include animals such as mammals,including, but not limited to, primates (e.g., humans), cows, sheep,goats, horses, dogs, cats, rabbits, rats, mice and the like. In someembodiments, the subject is a human.

As used herein, the term “sEH-mediated disease or condition” and thelike refers to a disease or condition characterized by less than orgreater than normal, sEH activity. A sEH-mediated disease or conditionis one in which modulation of sEH results in some effect on theunderlying condition or disease (e.g., a sEH inhibitor or antagonistresults in some improvement in patient well-being in at least somepatients).

“Parenchyma” refers to the tissue characteristic of an organ, asdistinguished from associated connective or supporting tissues.

“Chronic Obstructive Pulmonary Disease” or “COPD” is also sometimesknown as “chronic obstructive airway disease”, “chronic obstructive lungdisease”, and “chronic airways disease.” COPD is generally defined as adisorder characterized by reduced maximal expiratory flow and slowforced emptying of the lungs. COPD is considered to encompass tworelated conditions, emphysema and chronic bronchitis. COPD can bediagnosed by the general practitioner using art recognized techniques,such as the patient's forced vital capacity (“FVC”), the maximum volumeof air that can be forcibly expelled after a maximal inhalation. In theoffices of general practitioners, the FVC is typically approximated by a6 second maximal exhalation through a spirometer. The definition,diagnosis and treatment of COPD, emphysema, and chronic bronchitis arewell known in the art and discussed in detail by, for example, Honig andIngram, in Harrison's Principles of Internal Medicine, (Fauci et al.,Eds.), 14th Ed., 1998, McGraw-Hill, New York, pp. 1451-1460 (hereafter,“Harrison's Principles of Internal Medicine”).

“Emphysema” is a disease of the lungs characterized by permanentdestructive enlargement of the airspaces distal to the terminalbronchioles without obvious fibrosis.

“Chronic bronchitis” is a disease of the lungs characterized by chronicbronchial secretions which last for most days of a month, for threemonths a year, for two years.

As the names imply, “obstructive pulmonary disease” and “obstructivelung disease” refer to obstructive diseases, as opposed to restrictivediseases. These diseases particularly include COPD, bronchial asthma andsmall airway disease.

“Small airway disease.” There is a distinct minority of patients whoseairflow obstruction is due, solely or predominantly to involvement ofthe small airways. These are defined as airways less than 2 mm indiameter and correspond to small cartilaginous bronchi, terminalbronchioles and respiratory bronchioles. Small airway disease (SAD)represents luminal obstruction by inflammatory and fibrotic changes thatincrease airway resistance. The obstruction may be transient orpermanent.

The “interstitial lung diseases (ILDs)” are a group of conditionsinvolving the alveolar walls, perialveolar tissues, and contiguoussupporting structures. As discussed on the website of the American LungAssociation, the tissue between the air sacs of the lung is theinterstitium, and this is the tissue affected by fibrosis in thedisease. Persons with the disease have difficulty breathing in becauseof the stiffness of the lung tissue but, in contrast to persons withobstructive lung disease, have no difficulty breathing out. Thedefinition, diagnosis and treatment of interstitial lung diseases arewell known in the art and discussed in detail by, for example, Reynolds,H. Y., in Harrison's Principles of Internal Medicine, supra, at pp.1460-1466. Reynolds notes that, while ILDs have various initiatingevents, the immunopathological responses of lung tissue are limited andthe ILDs therefore have common features.

“Idiopathic pulmonary fibrosis,” or “IPF,” is considered the prototypeILD. Although it is idiopathic in that the cause is not known, Reynolds,supra, notes that the term refers to a well defined clinical entity.

“Bronchoalveolar lavage,” or “BAL,” is a test which permits removal andexamination of cells from the lower respiratory tract and is used inhumans as a diagnostic procedure for pulmonary disorders such as IPF. Inhuman patients, it is usually performed during bronchoscopy.

As used herein, the term “alkyl” refers to a saturated hydrocarbonradical which may be straight-chain or branched-chain (for example,ethyl, isopropyl, t-amyl, or 2,5-dimethylhexyl). This definition appliesboth when the term is used alone and when it is used as part of acompound term, such as “arylalkyl,” “alkylamino” and similar terms. Insome embodiments, alkyl groups are those containing 1 to 24 carbonatoms. All numerical ranges in this specification and claims areintended to be inclusive of their upper and lower limits. Additionally,the alkyl and heteroalkyl groups may be attached to other moieties atany position on the alkyl or heteroalkyl radical which would otherwisebe occupied by a hydrogen atom (such as, for example, 2-pentyl,2-methylpent-1-yl and 2-propyloxy). Divalent alkyl groups may bereferred to as “alkylene,” and divalent heteroalkyl groups may bereferred to as “heteroalkylene,” such as those groups used as linkers inthe present invention. The alkyl, alkylene, and heteroalkylene moietiesmay also be optionally substituted with halogen atoms, or other groupssuch as oxo, cyano, nitro, alkyl, alkylamino, carboxyl, hydroxyl,alkoxy, aryloxy, and the like.

The terms “cycloalkyl” and “cycloalkylene” refer to a saturatedhydrocarbon ring and includes bicyclic and polycyclic rings. Similarly,cycloalkyl and cycloalkylene groups having a heteroatom (e.g. N, O or S)in place of a carbon ring atom may be referred to as “heterocycloalkyl”and “heterocycloalkylene,” respectively. Examples of cycloalkyl andheterocycloalkyl groups are, for example, cyclohexyl, norbornyl,adamantyl, morpholinyl, thiomorpholinyl, dioxothiomorpholinyl, and thelike. The cycloalkyl and heterocycloalkyl moieties may also beoptionally substituted with halogen atoms, or other groups such asnitro, alkyl, alkylamino, carboxyl, alkoxy, aryloxy and the like. Insome embodiments, cycloalkyl and cycloalkylene moieties are those having3 to 12 carbon atoms in the ring (e.g., cyclohexyl, cyclooctyl,norbornyl, adamantyl, and the like). In some embodiments,heterocycloalkyl and heterocycloalkylene moieties are those having 1 to3 hetero atoms in the ring (e.g., morpholinyl, thiomorpholinyl,dioxothiomorpholinyl, piperidinyl and the like). Additionally, the term“(cycloalkyl)alkyl” refers to a group having a cycloalkyl moietyattached to an alkyl moiety. Examples are cyclohexylmethyl,cyclohexylethyl and cyclopentylpropyl.

The term “alkenyl” as used herein refers to an alkyl group as describedabove which contains one or more sites of unsaturation that is a doublebond. Similarly, the term “alkynyl” as used herein refers to an alkylgroup as described above which contains one or more sites ofunsaturation that is a triple bond.

The term “alkoxy” refers to an alkyl radical as described above whichalso bears an oxygen substituent which is capable of covalent attachmentto another hydrocarbon radical (such as, for example, methoxy, ethoxyand t-butoxy).

The term “aryl” refers to an aromatic carbocyclic substituent which maybe a single ring or multiple rings which are fused together, linkedcovalently or linked to a common group such as an ethylene or methylenemoiety. Similarly, aryl groups having a heteroatom (e.g. N, O or S) inplace of a carbon ring atom are referred to as “heteroaryl”. Examples ofaryl and heteroaryl groups are, for example, phenyl, naphthyl, biphenyl,diphenylmethyl, thienyl, pyridyl and quinoxalyl. The aryl and heteroarylmoieties may also be optionally substituted with halogen atoms, or othergroups such as nitro, alkyl, alkylamino, carboxyl, alkoxy, phenoxy andthe like. Additionally, the aryl and heteroaryl groups may be attachedto other moieties at any position on the aryl or heteroaryl radicalwhich would otherwise be occupied by a hydrogen atom (such as, forexample, 2-pyridyl, 3-pyridyl and 4-pyridyl). Divalent aryl groups are“arylene”, and divalent heteroaryl groups are referred to as“heteroarylene” such as those groups used as linkers in the presentinvention.

The terms “arylalkyl” and “alkylaryl”, “refer to an aryl radicalattached directly to an alkyl group. Likewise, the terms “arylalkenyl”and “aryloxyalkyl” refer to an alkenyl group, or an oxygen which isattached to an alkyl group, respectively. For brevity, aryl as part of acombined term as above, is meant to include heteroaryl as well. The term“aryloxy” refers to an aryl radical as described above which also bearsan oxygen substituent which is capable of covalent attachment to anotherradical (such as, for example, phenoxy, naphthyloxy, and pyridyloxy).

The terms “halo” or “halogen,” by themselves or as part of anothersubstituent, mean, unless otherwise stated, a fluorine, chlorine,bromine, or iodine atom. Additionally, terms such as “haloalkyl,” and“haloalkoxy” are meant to include monohaloalkyl(oxy) andpolyhaloalkyl(oxy). For example, the term “C₁-C₆ haloalkyl” is mean toinclude trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl,3-bromopropyl, and the like.

The term “hetero” as used in a “heteroatom-containing alkyl group” (a“heteroalkyl” group) or a “heteroatom-containing aryl group” (a“heteroaryl” group) refers to a molecule, linkage or substituent inwhich one or more carbon atoms are replaced with an atom other thancarbon, e.g., nitrogen, oxygen, sulfur, phosphorus or silicon, typicallynitrogen, oxygen or sulfur or more than one non-carbon atom (e.g.,sulfonamide). Similarly, the term “heteroalkyl” refers to an alkylsubstituent that is heteroatom-containing, the terms “heterocyclic”“heterocycle” or “heterocyclyl” refer to a cyclic substituent or groupthat is heteroatom-containing and is either aromatic or non-aromatic.The terms “heteroaryl” and “heteroaromatic” respectively refer to “aryl”and “aromatic” substituents that are heteroatom-containing, and thelike. The terms “heterocyclic” and “heterocyclyl” include the terms“heteroaryl” and “heteroaromatic”. In some embodiments, heterocyclicmoieties are those having 1 to 3 hetero atoms in the ring. Examples ofheteroalkyl groups include alkoxy, alkoxyaryl, alkylsulfanyl-substitutedalkyl, N-alkylated amino alkyl, and the like. Examples of heteroarylsubstituents include pyrrolyl, pyrrolidinyl, pyridinyl, quinolinyl,indolyl, pyrimidinyl, imidazolyl, 1,2,4-triazolyl, tetrazolyl, etc., andexamples of heteroatom-containing cyclic nonaromatic groups aremorpholinyl, piperazinyl, piperidinyl, etc.

The term “carboxylic acid analog” refers to a variety of groups havingan acidic moiety that are capable of mimicking a carboxylic acidresidue. Examples of such groups are sulfonic acids, sulfinic acids,phosphoric acids, phosphonic acids, phosphinic acids, sulfonamides, andheterocyclic moieties such as, for example, imidazoles, triazoles andtetrazoles.

The term “substituted” refers to the replacement of an atom or a groupof atoms of a compound with another atom or group of atoms. For example,an atom or a group of atoms may be substituted with one or more of thefollowing substituents or groups: halo, nitro, C₁-C₈alkyl,C₁-C₈alkylamino, hydroxyC₁-C₈alkyl, haloC₁-C₈alkyl, carboxyl, hydroxyl,C₁-C₈alkoxy, C₁-C₈alkoxyC₁-C₈alkoxy, thioC₁-C₈alkyl, aryl, aryloxy,C₃-C₈cycloalkyl, C₃-C₈cycloalkyl C₁-C₈alkyl, heteroaryl, arylC₁-C₈alkyl,heteroarylC₁-C₈alkyl, C₂-C₈alkenyl containing 1 to 2 double bonds,C₂-C₈alkynyl containing 1 to 2 triple bonds, C₄-C₈alk(en)(yn)yl groups,cyano, formyl, C₁-C₈alkylcarbonyl, arylcarbonyl, heteroarylcarbonyl,C₁-C₈alkoxycarbonyl, aryloxycarbonyl, aminocarbonyl,C₁-C₈alkylaminocarbonyl, C₁-C₈dialkylaminocarbonyl, arylaminocarbonyl,diarylaminocarbonyl, arylC₁-C₈alkylaminocarbonyl, haloC₁-C₈alkoxy,C₂-C₈alkenyloxy, C₂-C₈alkynyloxy, arylC₁-C₈alkoxy, aminoC₁-C₈alkyl,C₁-C₈alkylaminoC₁-C₈alkyl, C₁-C₈dialkylaminoC₁-C₈alkyl,arylaminoC₁-C₈alkyl, amino, C₁-C₈dialkylamino, arylamino,arylC₁-C₈alkylamino, C₁-C₈alkylcarbonylamino, arylcarbonylamino, azido,mercapto, C₁-C₈alkylthio, arylthio, haloC₁-C₈alkylthio, thiocyano,isothiocyano, C₁-C₈alkylsulfinyl, C₁-C₈alkylsulfonyl, arylsulfinyl,arylsulfonyl, aminosulfonyl, C₁-C₈alkylaminosulfonyl,C₁-C₈dialkylaminosulfonyl and arylaminosulfonyl. When the term“substituted” appears prior to a list of possible substituted groups, itis intended that the term apply to every member of that group.

The term “unsubstituted” refers to a native compound that lacksreplacement of an atom or a group of atoms.

General:

The present invention derives from the discovery that 1,3-disubstitutedureas (or the corresponding amides or carbamates, also referred to asthe primary pharmacophore) can be further functionalized to provide morepotent sEH inhibitors with improved physical properties. As describedherein, the introduction of a heterocyclic moiety can increase watersolubility and oral availability of sEH inhibitors (see below). Thecombination of these moieties provides a variety of compounds ofincreased water solubility.

The discovery of the heterocyclic pharmacophores has also led to theemployment of combinatorial chemistry approaches for establishing a widespectrum of compounds having sEH inhibitory activity. The polarpharmacophores divide the molecule into domains each of which can beeasily manipulated by common chemical approaches in a combinatorialmanner, leading to the design and confirmation of novel orally availabletherapeutic agents for the treatment of diseases such as hypertensionand vascular inflammation. The agents of the present invention treatsuch diseases while simultaneously increasing sodium excretion, reducingvascular and renal inflammation, and reducing male erectile dysfunctionAs shown below (see Examples and Figures), alterations in solubility,bioavailability and pharmacological properties leads to compounds thatcan alter the regulatory lipids of experimental animals increasing therelative amounts of epoxy arachidonate derivatives when compared eitherto their diol products or to the proinflammatory and hypertensivehydroxyeicosatetraenoic acids (HETEs). Since epoxy arachidonates areanti-hypertensive and anti-inflammatory, altering the lipid ratios canlead to reduced blood pressure and reduced vascular and renalinflammation. This approach has been validated as reported in U.S.patent application Ser. Nos. 10/817,334 and 11/256,685 which are hereinincorporated by reference in their entirety.

The heterocyclic group improves water solubility of sEH inhibitors aswell as the specificity for the sEH, and a wide diversity offunctionalities such as an ester, amide, carbamate, or similarfunctionalities capable of donating or accepting a hydrogen bondsimilarly can contribute to this polar group. For example, inpharmaceutical chemistry heterocyclic groups are commonly used to mimiccarbonyls as hydrogen bond donors and acceptors. Of course the primary,secondary and tertiary pharmacophore groups can be combined in a singlemolecule with suitable spacers to improve activity or present theinhibitor as a prodrug.

Methods of Inhibiting Soluble Epoxide Hydrolases:

In view of the above, the present invention provides, in one aspect, amethod for inhibiting a soluble epoxide hydrolase, comprising contactingthe soluble epoxide hydrolase with an inhibiting amount of a compoundhaving the formula (I):

The symbol R¹ is a member selected from the group consisting ofC₁-C₈alkyl, arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each ofwhich is optionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cyclic portions are monocyclic or polycyclic.In one embodiment, the 1 to 2 substituents are each independentlyselected from the group consisting of C₁-C₈alkyl and C₁-C₈alkoxy. In oneembodiment, the 1 to 2 substituents are each independently selected fromthe group consisting of C₁-C₈haloalkyl and C₁-C₈haloalkoxy.

The symbol Y¹ is selected from the group consisting of a bond, C(R⁵)₂,NR⁵ and O.

The symbol Y² is selected from the group consisting of a bond, NR⁵ andO.

Each symbol R², R³ and R⁵ is independently selected from the groupconsisting of H, C₁-C₈alkyl and COR⁶.

The symbol A is heterocyclyl, optionally substituted with from 1 to 2 R⁷substituents.

The symbol L is selected from the group consisting of a direct bond,C₁-C₁₂alkylene, C₁-C₁₂heteroalkylene, C₃-C₆cycloalkylene, arylene,heteroarylene, —CO—, −SO_(m)— and —Se—.

The symbol R⁴ is selected from the group consisting of H, C₁-C₈alkyl,C₂-C₆alkenyl, C₂-C₆alkynyl, C₁-C₈heteroalkyl, arylC₀-C₈alkyl,C₃-C₁₂cycloalkyl and heterocyclyl, each of which is optionallysubstituted. In one embodiment, each C₁-C₈alkyl, C₂-C₆alkenyl,C₂-C₆alkynyl, C₁-C₈heteroalkyl, arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl andheterocyclyl is optionally substituted with from 1 to 2 substituentseach independently selected from the group consisting of C₁-C₈alkyl,halo, C₁-C₈heteroalkyl, arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.In one embodiment, R⁴ is selected from the group consisting ofC₁-C₈alkyl and C₁-C₈alkoxy. In one embodiment, R⁴ is selected from thegroup consisting of C₁-C₈haloalkyl and C₁-C₈haloalkoxy.

Each symbol R⁶ is independently selected from the group consisting of H,C₁-C₈alkyl, OH, C₁-C₈alkoxy and amino.

Each symbol R⁷ is selected from the group consisting of halo, nitro,C₁-C₈alkyl, C₁-C₈alkylamino, hydroxyC₁-C₈alkyl, haloC₁-C₈alkyl,carboxyl, hydroxyl, C₁-C₈alkoxy, C₁-C₈alkoxyC₁-C₈alkoxy,haloC₁-C₈alkoxy, thioC₁-C₈alkyl, aryl, aryloxy, C₃-C₈cycloalkyl,C₃-C₈cycloalkyl C₁-C₈alkyl, heteroaryl, arylC₁-C₈alkyl,heteroarylC₁-C₈alkyl, C₂-C₈alkenyl containing 1 to 2 double bonds,C₂-C₈alkynyl containing 1 to 2 triple bonds, C₄-C₈alk(en)(yn)yl groups,cyano, formyl, C₁-C₈alkylcarbonyl, arylcarbonyl, heteroarylcarbonyl,C₁-C₈alkoxycarbonyl, aryloxycarbonyl, aminocarbonyl,C₁-C₈alkylaminocarbonyl, C₁-C₈dialkylaminocarbonyl, arylaminocarbonyl,diarylaminocarbonyl, arylC₁-C₈alkylaminocarbonyl, haloC₁-C₈alkoxy,C₂-C₈alkenyloxy, C₂-C₈alkynyloxy, arylC₁-C₈alkoxy, aminoC₁-C₈alkyl,C₁-C₈alkylaminoC₁-C₈alkyl, C₁-C₈dialkylaminoC₁-C₈alkyl,arylaminoC₁-C₈alkyl, amino, C₁-C₈dialkylamino, arylamino,arylC₁-C₈alkylamino, C₁-C₈alkylcarbonylamino, arylcarbonylamino, azido,mercapto, C₁-C₈alkylthio, arylthio, haloC₁-C₈alkylthio, thiocyano,isothiocyano, C₁-C₈alkylsulfinyl, C₁-C₈alkylsulfonyl, arylsulfinyl,arylsulfonyl, aminosulfonyl, C₁-C₈alkylaminosulfonyl,C₁-C₈dialkylaminosulfonyl and arylaminosulfonyl.

The subscript n is an integer of 0 to 1.

The subscript m is an integer of from 0 to 2.

The compounds include all pharmaceutically acceptable derivativesthereof, such as salts, prodrugs, solvates and hydrates.

In other embodiments Y¹ is NR⁵. In further embodiments Y² is a bond. Instill further embodiments Y² is NR⁵. In still other embodiments Y² is O.

In other embodiments Y² is NR⁵. In further embodiments Y¹ is a bond. Instill other embodiments Y¹ is C(R⁵)₂. In further embodiments Y¹ is O. Instill further embodiments Y¹ is NR⁵.

In other embodiments R², R³ and R⁵ are H.

In further embodiments, A is selected from the group consisting ofpiperidinyl, 1,3,5-triaza-tricyclo[3.3.1.13,7]decyl, indolyl, pyridyl,morpholinyl and benzimidazolyl. In still other embodiments A ispiperidinyl. In other embodiments A is1,3,5-triaza-tricyclo[3.3.1.13,7]decyl. In still further embodiments Ais indolyl. In other embodiments A is pyridyl. In other embodiments A ismorpholinyl. In other embodiments A is benzimidazolyl.

In still other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which areoptionally substituted. In further embodiments, each of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl are optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

the subscript n is an integer of 0 to 1; and

the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl. In one embodiment, R⁴is selected from the group consisting of C₁-C₈alkyl and C₁-C₈alkoxy. Inone embodiment, R⁴ is selected from the group consisting ofC₁-C₈haloalkyl and C₁-C₈haloalkoxy.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl;

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In other embodiments the compound has the formula:

wherein R¹ is a member selected from the group consisting of C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, C₁-C₈heteroalkyl, aryl,heteroaryl; wherein said cycloalkyl portions are monocyclic orpolycyclic.

Within these embodiments, L is selected from the group consisting of adirect bond, C₁-C₁₂heteroalkylene, —CO— and —SO_(m)—; and

R⁴ is selected from the group consisting of H, C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl, each of which isoptionally substituted. In one embodiment, each C₁-C₈alkyl,arylC₀-C₈alkyl, C₃-C₁₂cycloalkyl and heterocyclyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, halo, C₁-C₈heteroalkyl,arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl.

Within these embodiments, each R⁶ is independently selected from thegroup consisting of H, C₁-C₈alkyl, C₁-C₈alkoxy and amino;

-   the subscript n is an integer of 0 to 1; and-   the subscript m is an integer of from 0 to 2.

In any of the above embodiments R¹ is C₁-C₈alkyl. In any of the aboveembodiments R¹ is selected from the group consisting of dodecyl andt-butyl. In any of the above embodiments R¹ is arylC₀-C₈alkyl. In any ofthe above embodiments R¹ is phenyl. In any of the above embodiments R¹is C₃-C₁₂cycloalkyl. In any of the above embodiments R¹ is adamantyl. Inany of the above embodiments R¹ is cycloheptyl or cyclohexyl. In any ofthe above embodiments R¹ is C₃-C₁₂cycloalkyl. In any of the aboveembodiments R¹ is adamantyl. In any of the above embodiments R¹ iscycloheptyl. In any of the above embodiments of R¹, the group isoptionally substituted. In any of the above embodiments, the R¹ group isoptionally substituted with from 1 to 2 substituents. In any of theabove embodiments the 1 to 2 substituents are each independentlyselected from the group consisting of C₁-C₈alkyl and C₁-C₈alkoxy. In anyof the above embodiments embodiment, the 1 to 2 substituents are eachindependently selected from the group consisting of C₁-C₈haloalkyl andC₁-C₈haloalkoxy.

In any of the above embodiments L is a direct bond. In any of the aboveembodiments L is C₁-C₁₂heteroalkylene. In any of the above embodiments Lis —CO—. In any of the above embodiments L is —SO₂—.

In any of the above embodiments R⁴ is selected from the group consistingof H, C₁-C₈alkyl, arylC₀-C₈alkyl, C₁-C₈alkoxy and heterocyclyl. In anyof the above embodiments, R⁴ is selected from the group consisting ofC₁-C₈haloalkyl and C₁-C₈haloalkoxy.

In any of the above embodiments R⁶ is H. In any of the above embodimentsR⁶ is C₁-C₈alkyl.

In any of the above embodiments n is 0. In any of the above embodimentsn is 1.

In other embodiments, the compound is selected from the group consistingof the compounds of Examples 1-70 and Tables 1-4 and 5a and 5b.

In any of the above embodiments the compounds include allpharmaceutically acceptable derivatives thereof, such as salts,prodrugs, solvates and hydrates.

Assays to Monitor Soluble Epoxide Hydrolase Activity:

Additionally, the present invention provides a variety of assays andassociated methods for monitoring soluble epoxide hydrolase activity,particularly the activity that has been modulated by the administrationof one or more of the compounds provided above.

In one group of embodiments, the invention provides methods for reducingthe formation of a biologically active diol produced by the action of asoluble epoxide hydrolase, the method comprising contacting the solubleepoxide hydrolase with an amount of a compound of formula (I) above,sufficient to inhibit the activity of the soluble epoxide hydrolase andreduce the formation of the biologically active diol.

In another group of embodiments, the invention provides methods forstabilizing biologically active epoxides in the presence of a solubleepoxide hydrolase, the method comprising contacting the soluble epoxidehydrolase with an amount of a compound of formula (I), sufficient toinhibit the activity of the soluble epoxide hydrolase and stabilize thebiologically active epoxide.

In each of these groups of embodiments, the methods can be carried outas part of an in vitro assay or the methods can be carried out in vivoby monitoring blood titers of the respective biologically active epoxideor diol.

Epoxides and diols of some fatty acids are biologically importantchemical mediators and are involved in several biological processes. Thestrongest biological data support the action of oxylipins as chemicalmediators between the vascular endothelium and vascular smooth muscle.Epoxy lipids are anti-inflammatory and anti-hypertensive. Additionally,the lipids are thought to be metabolized by beta-oxidation, as well asby epoxide hydration. The soluble epoxide hydrolase is considered to bethe major enzyme involved in the hydrolytic metabolism of theseoxylipins. The compounds of formula (I) can inhibit the epoxidehydrolase and stabilize the epoxy lipids both in vitro and in vivo. Thisactivity results in a reduction of hypertension in four separate rodentmodels. Moreover, the inhibitors show a reduction in renal inflammationassociated with and independent of the hypertensive models.

More particularly, the present invention provides methods for monitoringa variety of lipids in both the arachidonate and linoleate cascadesimultaneously in order to address the biology of the system. A GLC-MSsystem or a LC-MS method can be used to monitor over 740 analytes in ahighly quantitative fashion in a single injection. The analytes includethe regioisomers of the arachidonate epoxides (EETs), the diols (DHETs),as well as other P450 products including HETEs. Characteristic productsof the cyclooxygenase, lipoxygenase, and peroxidase pathways in both thearachidonate and linoleate series can also be monitored. Such methodsare particularly useful as being predictive of certain disease states.The oxylipins can be monitored in mammals following the administrationof inhibitors of epoxide hydrolase. Generally, EH inhibitors increaseepoxy lipid concentrations at the expense of diol concentrations in bodyfluids and tissues.

Other compounds for use in this aspect of the invention are thoseinhibitors of formula (I) in which the primary pharmacophore isseparated from a secondary and/or tertiary pharmacophore by a distancethat approximates the distance between the terminal carboxylic acid andan epoxide functional group in the natural substrate.

Methods of Treating Diseases Modulated by Soluble Epoxide Hydrolases:

In another aspect, the present invention provides methods of treatingdiseases, especially those modulated by soluble epoxide hydrolases(sEH). The methods generally involve administering to a subject in needof such treatment an effective amount of a compound having a formula (I)above. The dose, frequency and timing of such administering will dependin large part on the selected therapeutic agent, the nature of thecondition being treated, the condition of the subject including age,weight and presence of other conditions or disorders, the formulationbeing administered and the discretion of the attending physician.Preferably, the compositions and compounds of the invention and thepharmaceutically acceptable salts thereof are administered via oral,parenteral, subcutaneous, intramuscular, intravenous or topical routes.Generally, the compounds are administered in dosages ranging from about2 mg up to about 2,000 mg per day, although variations will necessarilyoccur depending, as noted above, on the disease target, the patient, andthe route of administration. Dosages are administered orally in therange of about 0.05 mg/kg to about 20 mg/kg, more preferably in therange of about 0.05 mg/kg to about 2 mg/kg, most preferably in the rangeof about 0.05 mg/kg to about 0.2 mg per kg of body weight per day. Thedosage employed for the topical administration will, of course, dependon the size of the area being treated.

It has previously been shown that inhibitors of soluble epoxidehydrolase (“sEH”) can reduce hypertension. See, e.g., U.S. Pat. No.6,351,506. Such inhibitors can be useful in controlling the bloodpressure of persons with undesirably high blood pressure, includingthose who suffer from diabetes.

In some embodiments, compounds of formula (I) are administered to asubject in need of treatment for hypertension, specifically renal,hepatic, or pulmonary hypertension; inflammation, specifically renalinflammation, vascular inflammation, and lung inflammation; adultrespiratory distress syndrome; diabetic complications; end stage renaldisease; Raynaud syndrome and arthritis.

Methods for Inhibiting Progression of Kidney Deterioration (Nephropathy)and Reducing Blood Pressure:

In another aspect of the invention, the compounds of the invention canreduce damage to the kidney, and especially damage to kidneys fromdiabetes, as measured by albuminuria. The compounds of the invention canreduce kidney deterioration (nephropathy) from diabetes even inindividuals who do not have high blood pressure. The conditions oftherapeautic administration are as described above.

cis-Epoxyeicosantrienoic acids (“EETs”) can be used in conjunction withthe compounds of the invention to further reduce kidney damage. EETs,which are epoxides of arachidonic acid, are known to be effectors ofblood pressure, regulators of inflammation, and modulators of vascularpermeability. Hydrolysis of the epoxides by sEH diminishes thisactivity. Inhibition of sEH raises the level of EETs since the rate atwhich the EETs are hydrolyzed into DHETs is reduced. Without wishing tobe bound by theory, it is believed that raising the level of EETsinterferes with damage to kidney cells by the microvasculature changesand other pathologic effects of diabetic hyperglycemia. Therefore,raising the EET level in the kidney is believed to protect the kidneyfrom progression from microalbuminuria to end stage renal disease.

EETs are well known in the art. EETs useful in the methods of thepresent invention include 14,15-EET, 8,9-EET and 11,12-EET, and 5,6EETs, in that order of preference. Preferably, the EETs are administeredas the methyl ester, which is more stable. Persons of skill willrecognize that the EETs are regioisomers, such as 8S,9R- and14R,15S-EET. 8,9-EET, 11,12-EET, and 14R,15S-EET, are commerciallyavailable from, for example, Sigma-Aldrich (catalog nos. E5516, E5641,and E5766, respectively, Sigma-Aldrich Corp., St. Louis, Mo.).

EETs produced by the endothelium have anti-hypertensive properties andthe EETs 11,12-EET and 14,15-EET may be endothelium-derivedhyperpolarizing factors (EDHFs). Additionally, EETs such as 11,12-EEThave profibrinolytic effects, anti-inflammatory actions and inhibitsmooth muscle cell proliferation and migration. In the context of thepresent invention, these favorable properties are believed to protectthe vasculature and organs during renal and cardiovascular diseasestates.

It is now believed that sEH activity can be inhibited sufficiently toincrease the levels of EETs and thus augment the effects ofadministering sEH inhibitors by themselves. This permits EETs to be usedin conjunction with one or more sEH inhibitors to reduce nephropathy inthe methods of the invention. It further permits EETs to be used inconjunction with one or more sEH inhibitors to reduce hypertension, orinflammation, or both. Thus, medicaments of EETs can be made which canbe administered in conjunction with one or more sEH inhibitors, or amedicament containing one or more sEH inhibitors can optionally containone or more EETs.

The EETs can be administered concurrently with the sEH inhibitor, orfollowing administration of the sEH inhibitor. It is understood that,like all drugs, inhibitors have half lives defined by the rate at whichthey are metabolized by or excreted from the body, and that theinhibitor will have a period following administration during which itwill be present in amounts sufficient to be effective. If EETs areadministered after the inhibitor is administered, therefore, it isdesirable that the EETs be administered during the period during whichthe inhibitor will be present in amounts to be effective to delayhydrolysis of the EETs. Typically, the EET or EETs will be administeredwithin 48 hours of administering an sEH inhibitor. Preferably, the EETor EETs are administered within 24 hours of the inhibitor, and even morepreferably within 12 hours. In increasing order of desirability, the EETor EETs are administered within 10, 8, 6, 4, 2, hours, 1 hour, or onehalf hour after administration of the inhibitor. Most preferably, theEET or EETs are administered concurrently with the inhibitor.

In some embodiments, the EETs, the compound of the invention, or both,are provided in a material that permits them to be released over time toprovide a longer duration of action. Slow release coatings are wellknown in the pharmaceutical art; the choice of the particular slowrelease coating is not critical to the practice of the presentinvention.

EETs are subject to degradation under acidic conditions. Thus, if theEETs are to be administered orally, it is desirable that they areprotected from degradation in the stomach. Conveniently, EETs for oraladministration may be coated to permit them to passage the acidicenvironment of the stomach into the basic environment of the intestines.Such coatings are well known in the art. For example, aspirin coatedwith so-called “enteric coatings” is widely available commercially. Suchenteric coatings may be used to protect EETs during passage through thestomach. An exemplary coating is set forth in the Examples.

While the anti-hypertensive effects of EETs have been recognized, EETshave not been administered to treat hypertension because it was thoughtendogenous sEH would hydrolyse the EETs too quickly for them to have anyuseful effect. Surprisingly, it was found during the course of thestudies underlying the present invention that exogenously administeredinhibitors of sEH succeeded in inhibiting sEH sufficiently that levelsof EETs could be further raised by the administration of exogenous EETs.These findings underlie the co-administration of sEH inhibitors and ofEETs described above with respect to inhibiting the development andprogression of nephropathy. This is an important improvement inaugmenting treatment. While levels of endogenous EETs are expected torise with the inhibition of sEH activity caused by the action of the sEHinhibitor, and therefore to result in at least some improvement insymptoms or pathology, it may not be sufficient in all cases to inhibitprogression of kidney damage fully or to the extent intended. This isparticularly true where the diseases or other factors have reduced theendogenous concentrations of EETs below those normally present inhealthy individuals. Administration of exogenous EETs in conjunctionwith a sEH inhibitor is therefore expected to be beneficial and toaugment the effects of the sEH inhibitor in reducing the progression ofdiabetic nephropathy.

The present invention can be used with regard to any and all forms ofdiabetes to the extent that they are associated with progressive damageto the kidney or kidney function. The chronic hyperglycemia of diabetesis associated with long-term damage, dysfunction, and failure of variousorgans, especially the eyes, kidneys, nerves, heart, and blood vessels.The long-term complications of diabetes include retinopathy withpotential loss of vision; nephropathy leading to renal failure;peripheral neuropathy with risk of foot ulcers, amputation, and Charcotjoints.

In addition, persons with metabolic syndrome are at high risk ofprogression to type 2 diabetes, and therefore at higher risk thanaverage for diabetic nephropathy. It is therefore desirable to monitorsuch individuals for microalbuminuria, and to administer a sEH inhibitorand, optionally, one or more EETs, as an intervention to reduce thedevelopment of nephropathy. The practitioner may wait untilmicroalbuminuria is seen before beginning the intervention. As notedabove, a person can be diagnosed with metabolic syndrome without havinga blood pressure of 130/85 or higher. Both persons with blood pressureof 130/85 or higher and persons with blood pressure below 130/85 canbenefit from the administration of sEH inhibitors and, optionally, ofone or more EETs, to slow the progression of damage to their kidneys. Insome embodiments, the person has metabolic syndrome and blood pressurebelow 130/85.

Dyslipidemia or disorders of lipid metabolism is another risk factor forheart disease. Such disorders include an increased level of LDLcholesterol, a reduced level of HDL cholesterol, and an increased levelof triglycerides. An increased level of serum cholesterol, andespecially of LDL cholesterol, is associated with an increased risk ofheart disease. The kidneys are also damaged by such high levels. It isbelieved that high levels of triglycerides are associated with kidneydamage. In particular, levels of cholesterol over 200 mg/dL, andespecially levels over 225 mg/dL, would suggest that sEH inhibitors and,optionally, EETs, should be administered. Similarly, triglyceride levelsof more than 215 mg/dL, and especially of 250 mg/dL or higher, wouldindicate that administration of sEH inhibitors and, optionally, of EETs,would be desirable. The administration of compounds of the presentinvention with or without the EETs, can reduce the need to administerstatin drugs (HMG-CoA reductase inhibitors) to the patients, or reducethe amount of the statins needed. In some embodiments, candidates forthe methods, uses and compositions of the invention have triglyceridelevels over 215 mg/dL and blood pressure below 130/85. In someembodiments, the candidates have triglyceride levels over 250 mg/dL andblood pressure below 130/85. In some embodiments, candidates for themethods, uses and compositions of the invention have cholesterol levelsover 200 mg/dL and blood pressure below 130/85. In some embodiments, thecandidates have cholesterol levels over 225 mg/dL and blood pressurebelow 130/85.

Methods of Inhibiting the Proliferation of Vascular Smooth Muscle Cells:

In other embodiments, compounds of formula (I) inhibit proliferation ofvascular smooth muscle (VSM) cells without significant cell toxicity,(e.g., specific to VSM cells). Because VSM cell proliferation is anintegral process in the pathophysiology of atherosclerosis, thesecompounds are suitable for slowing or inhibiting atherosclerosis. Thesecompounds are useful to subjects at risk for atherosclerosis, such asindividuals who have had a heart attack or a test result showingdecreased blood circulation to the heart. The conditions of therapeuticadministration are as described above.

The methods of the invention are particularly useful for patients whohave had percutaneous intervention, such as angioplasty to reopen anarrowed artery, to reduce or to slow the narrowing of the reopenedpassage by restenosis. In some embodiments, the artery is a coronaryartery. The compounds of the invention can be placed on stents inpolymeric coatings to provide a controlled localized release to reducerestenosis. Polymer compositions for implantable medical devices, suchas stents, and methods for embedding agents in the polymer forcontrolled release, are known in the art and taught, for example, inU.S. Pat. Nos. 6,335,029; 6,322,847; 6,299,604; 6,290,722; 6,287,285;and 5,637,113. In some embodiments, the coating releases the inhibitorover a period of time, preferably over a period of days, weeks, ormonths. The particular polymer or other coating chosen is not a criticalpart of the present invention.

The methods of the invention are useful for slowing or inhibiting thestenosis or restenosis of natural and synthetic vascular grafts. Asnoted above in connection with stents, desirably, the synthetic vasculargraft comprises a material which releases a compound of the inventionover time to slow or inhibit VSM proliferation and the consequentstenosis of the graft. Hemodialysis grafts are a particular embodiment.

In addition to these uses, the methods of the invention can be used toslow or to inhibit stenosis or restenosis of blood vessels of personswho have had a heart attack, or whose test results indicate that theyare at risk of a heart attack.

In one group of embodiments, compounds of the invention are administeredto reduce proliferation of VSM cells in persons who do not havehypertension. In another group of embodiments, compounds of theinvention are used to reduce proliferation of VSM cells in persons whoare being treated for hypertension, but with an agent that is not an sEHinhibitor.

The compounds of the invention can be used to interfere with theproliferation of cells which exhibit inappropriate cell cycleregulation. In one important set of embodiments, the cells are cells ofa cancer. The proliferation of such cells can be slowed or inhibited bycontacting the cells with a compound of the invention. The determinationof whether a particular compound of the invention can slow or inhibitthe proliferation of cells of any particular type of cancer can bedetermined using assays routine in the art.

In addition to the use of the compounds of the invention, the levels ofEETs can be raised by adding EETs. VSM cells contacted with both an EETand a compound of the invention exhibited slower proliferation thancells exposed to either the EET alone or to the a compound of theinvention alone. Accordingly, if desired, the slowing or inhibition ofVSM cells of a compound of the invention can be enhanced by adding anEET along with a compound of the invention. In the case of stents orvascular grafts, for example, this can conveniently be accomplished byembedding the EET in a coating along with a compound of the invention sothat both are released once the stent or graft is in position.

Methods of Inhibiting the Progression of Obstructive Pulmonary Disease,Interstitial Lung Disease, or Asthma:

Chronic obstructive pulmonary disease, or COPD, encompasses twoconditions, emphysema and chronic bronchitis, which relate to damagecaused to the lung by air pollution, chronic exposure to chemicals, andtobacco smoke. Emphysema as a disease relates to damage to the alveoliof the lung, which results in loss of the separation between alveoli anda consequent reduction in the overall surface area available for gasexchange. Chronic bronchitis relates to irritation of the bronchioles,resulting in excess production of mucin, and the consequent blocking bymucin of the airways leading to the alveoli. While persons withemphysema do not necessarily have chronic bronchitis or vice versa, itis common for persons with one of the conditions to also have the other,as well as other lung disorders.

Some of the damage to the lungs due to COPD, emphysema, chronicbronchitis, and other obstructive lung disorders can be inhibited orreversed by administering inhibitors of the enzyme known as solubleepoxide hydrolase, or “sEH”. The effects of sEH inhibitors can beincreased by also administering EETs. The effect is at least additiveover administering the two agents separately, and may indeed besynergistic.

The studies reported herein show that EETs can be used in conjunctionwith sEH inhibitors to reduce damage to the lungs by tobacco smoke or,by extension, by occupational or environmental irritants. These findingsindicate that the co-administration of sEH inhibitors and of EETs can beused to inhibit or slow the development or progression of COPD,emphysema, chronic bronchitis, or other chronic obstructive lungdiseases which cause irritation to the lungs.

Animal models of COPD and humans with COPD have elevated levels ofimmunomodulatory lymphocytes and neutrophils. Neutrophils release agentsthat cause tissue damage and, if not regulated, will over time have adestructive effect. Without wishing to be bound by theory, it isbelieved that reducing levels of neutrophils reduces tissue damagecontributing to obstructive lung diseases such as COPD, emphysema, andchronic bronchitis. Administration of sEH inhibitors to rats in ananimal model of COPD resulted in a reduction in the number ofneutrophils found in the lungs. Administration of EETs in addition tothe sEH inhibitors also reduced neutrophil levels. The reduction inneutrophil levels in the presence of sEH inhibitor and EETs was greaterthan in the presence of the sEH inhibitor alone.

While levels of endogenous EETs are expected to rise with the inhibitionof sEH activity caused by the action of the sEH inhibitor, and thereforeto result in at least some improvement in symptoms or pathology, it maynot be sufficient in all cases to inhibit progression of COPD or otherpulmonary diseases. This is particularly true where the diseases orother factors have reduced the endogenous concentrations of EETs belowthose normally present in healthy individuals. Administration ofexogenous EETs in conjunction with an sEH inhibitor is thereforeexpected to augment the effects of the sEH inhibitor in inhibiting orreducing the progression of COPD or other pulmonary diseases.

In addition to inhibiting or reducing the progression of chronicobstructive airway conditions, the invention also provides new ways ofreducing the severity or progression of chronic restrictive airwaydiseases. While obstructive airway diseases tend to result from thedestruction of the lung parenchyma, and especially of the alveoli,restrictive diseases tend to arise from the deposition of excesscollagen in the parenchyma. These restrictive diseases are commonlyreferred to as “interstitial lung diseases”, or “ILDs”, and includeconditions such as idiopathic pulmonary fibrosis. The methods,compositions and uses of the invention are useful for reducing theseverity or progression of ILDs, such as idiopathic pulmonary fibrosis.Macrophages play a significant role in stimulating interstitial cells,particularly fibroblasts, to lay down collagen. Without wishing to bebound by theory, it is believed that neutrophils are involved inactivating macrophages, and that the reduction of neutrophil levelsfound in the studies reported herein demonstrate that the methods anduses of the invention will also be applicable to reducing the severityand progression of ILDs.

In some embodiments, the ILD is idiopathic pulmonary fibrosis. In otherembodiments, the ILD is one associated with an occupational orenvironmental exposure. Exemplars of such ILDs, are asbestosis,silicosis, coal worker's pneumoconiosis, and berylliosis. Further,occupational exposure to any of a number of inorganic dusts and organicdusts is believed to be associated with mucus hypersecretion andrespiratory disease, including cement dust, coke oven emissions, mica,rock dusts, cotton dust, and grain dust (for a more complete list ofoccupational dusts associated with these conditions, see Table 254-1 ofSpeizer, “Environmental Lung Diseases,” Harrison's Principles ofInternal Medicine, infra, at pp. 1429-1436). In other embodiments, theILD is sarcoidosis of the lungs. ILDs can also result from radiation inmedical treatment, particularly for breast cancer, and from connectivetissue or collagen diseases such as rheumatoid arthritis and systemicsclerosis. It is believed that the methods, uses and compositions of theinvention can be useful in each of these interstitial lung diseases.

In another set of embodiments, the invention is used to reduce theseverity or progression of asthma. Asthma typically results in mucinhypersecretion, resulting in partial airway obstruction. Additionally,irritation of the airway results in the release of mediators whichresult in airway obstruction. While the lymphocytes and otherimmunomodulatory cells recruited to the lungs in asthma may differ fromthose recruited as a result of COPD or an ILD, it is expected that theinvention will reduce the influx of immunomodulatory cells, such asneutrophils and eosinophils, and ameliorate the extent of obstruction.Thus, it is expected that the administration of sEH inhibitors, and theadministration of sEH inhibitors in combination with EETs, will beuseful in reducing airway obstruction due to asthma.

In each of these diseases and conditions, it is believed that at leastsome of the damage to the lungs is due to agents released by neutrophilswhich infiltrate into the lungs. The presence of neutrophils in theairways is thus indicative of continuing damage from the disease orcondition, while a reduction in the number of neutrophils is indicativeof reduced damage or disease progression. Thus, a reduction in thenumber of neutrophils in the airways in the presence of an agent is amarker that the agent is reducing damage due to the disease orcondition, and is slowing the further development of the disease orcondition. The number of neutrophils present in the lungs can bedetermined by, for example, bronchoalveolar lavage.

Prophylatic and Therapeutic Methods to Reduce Stroke Damage

Inhibitors of soluble epoxide hydrolase (“sEH”) and EETs administered inconjunction with inhibitors of sEH have been shown to reduce braindamage from strokes. Based on these results, we expect that inhibitorsof sEH taken prior to an ischemic stroke will reduce the area of braindamage and will likely reduce the consequent degree of impairment. Thereduced area of damage should also be associated with a faster recoveryfrom the effects of the stroke.

While the pathophysiologies of different subtypes of stroke differ, theyall cause brain damage. Hemorrhagic stroke differs from ischemic strokein that the damage is largely due to compression of tissue as bloodbuilds up in the confined space within the skull after a blood vesselruptures, whereas in ischemic stroke, the damage is largely due to lossof oxygen supply to tissues downstream of the blockage of a blood vesselby a clot. Ischemic strokes are divided into thrombotic strokes, inwhich a clot blocks a blood vessel in the brain, and embolic strokes, inwhich a clot formed elsewhere in the body is carried through the bloodstream and blocks a vessel there. But, in both hemorrhagic stroke andischemic stroke, the damage is due to the death of brain cells. Based onthe results observed in our studies, however, we would expect at leastsome reduction in brain damage in all types of stroke and in allsubtypes.

A number of factors are associated with an increased risk of stroke.Given the results of the studies underlying the present invention, sEHinhibitors administered to persons with any one or more of the followingconditions or risk factors:high blood pressure, tobacco use, diabetes,carotid artery disease, peripheral artery disease, atrial fibrillation,transient ischemic attacks (TIAs), blood disorders such as high redblood cell counts and sickle cell disease, high blood cholesterol,obesity, alcohol use of more than one drink a day for women or twodrinks a day for men, use of cocaine, a family history of stroke, aprevious stroke or heart attack, or being elderly, will reduce the areaof brain damaged of a stroke. With respect to being elderly, the risk ofstroke increases for every 10 years. Thus, as an individual reaches 60,70, or 80, administration of sEH inhibitors has an increasingly largerpotential benefit. As noted in the next section, the administration ofEETs in combination with one or more sEH inhibitors can be beneficial infurther reducing the brain damage. One can expect beneficial effectsfrom sEHI with or without EETs in a variety of diseases which lead toischemia reperfusion injury such as heart attacks.

In some uses and methods, the sEH inhibitors and, optionally, EETs, areadministered to persons who use tobacco, have carotid artery disease,have peripheral artery disease, have atrial fibrillation, have had oneor more transient ischemic attacks (TIAs), have a blood disorder such asa high red blood cell count or sickle cell disease, have high bloodcholesterol, are obese, use alcohol in excess of one drink a day if awoman or two drinks a day if a man, use cocaine, have a family historyof stroke, have had a previous stroke or heart attack and do not havehigh blood pressure or diabetes, or are 60, 70, or 80 years of age ormore and do not have hypertension or diabetes.

Clot dissolving agents, such as tissue plasminogen activator (tPA), havebeen shown to reduce the extent of damage from ischemic strokes ifadministered in the hours shortly after a stroke. tPA, for example, isapproved by the FDA for use in the first three hours after a stroke.Thus, at least some of the brain damage from a stroke is notinstantaneous, but occurs over a period of time or after a period oftime has elapsed after the stroke. It is therefore believed thatadministration of sEH inhibitors, optionally with EETs, can also reducebrain damage if administered within 6 hours after a stroke has occurred,more preferably within 5, 4, 3, or 2 hours after a stroke has occurred,with each successive shorter interval being more preferable. Even morepreferably, the inhibitor or inhibitors are administered 2 hours or lessor even 1 hour or less after the stroke, to maximize the reduction inbrain damage. Persons of skill are well aware of how to make a diagnosisof whether or not a patient has had a stroke. Such determinations aretypically made in hospital emergency rooms, following standarddifferential diagnosis protocols and imaging procedures.

In some uses and methods, the sEH inhibitors and, optionally, EETs, areadministered to persons who have had a stroke within the last 6 hourswho: use tobacco, have carotid artery disease, have peripheral arterydisease, have atrial fibrillation, have had one or more transientischemic attacks (TIAs), have a blood disorder such as a high red bloodcell count or sickle cell disease, have high blood cholesterol, areobese, use alcohol in excess of one drink a day if a woman or two drinksa day if a man, use cocaine, have a family history of stroke, have had aprevious stroke or heart attack and do not have high blood pressure ordiabetes, or are 60, 70, or 80 years of age or more and do not havehypertension or diabetes.

The conditions of therapeautic administration for all of theseindications are as described above.

Combination Therapy

As noted above, the compounds of the present invention will, in someinstances, be used in combination with other therapeutic agents to bringabout a desired effect. Selection of additional agents will, in largepart, depend on the desired target therapy (see, e.g., Turner, N. et al.Prog. Drug Res. (1998) 51: 33-94; Haffner, S. Diabetes Care (1998) 21:160-178; and DeFronzo, R. et al. (eds.), Diabetes Reviews (1997) Vol. 5No. 4). A number of studies have investigated the benefits ofcombination therapies with oral agents (see, e.g., Mahler, R., J. Clin.Endocrinol. Metab. (1999) 84: 1165-71; United Kingdom ProspectiveDiabetes Study Group: UKPDS 28, Diabetes Care (1998) 21: 87-92; Bardin,C. W., (ed.), Current Therapy In Endocrinology And Metabolism, 6thEdition (Mosby—Year Book, Inc., St. Louis, Mo. 1997); Chiasson, J. etal., Ann. Intern. Med. (1994) 121: 928-935; Coniff, R. et al., Clin.Ther. (1997) 19: 16-26; Coniff, R. et al., Am. J. Med. (1995) 98:443-451; and Iwamoto, Y. et al., Diabet. Med. (1996) 13 365-370;Kwiterovich, P. Am. J. Cardiol (1998) 82(12A): 3U-17U). Combinationtherapy includes administration of a single pharmaceutical dosageformulation which contains a compound having the general structure offormula I and one or more additional active agents, as well asadministration of a compound of formula I and each active agent in itsown separate pharmaceutical dosage formulation. For example, a compoundof formula I and one or more angiotensin receptor blockers, angiotensinconverting enzyme inhibitors, calcium channel blockers, diuretics, alphablockers, beta blockers, centrally acting agents, vasopeptidaseinhibitors, renin inhibitors, endothelin receptor agonists, AGEcrosslink breakers, sodium/potassium ATPase inhibitors, endothelinreceptor agonists, endothelin receptor antagonists, angiotensin vaccine,and the like; can be administered to the human subject together in asingle oral dosage composition, such as a tablet or capsule, or eachagent can be administered in separate oral dosage formulations. Whereseparate dosage formulations are used, a compound of formula I and oneor more additional active agents can be administered at essentially thesame time (i.e., concurrently), or at separately staggered times (i.e.,sequentially). Combination therapy is understood to include all theseregimens.

Compounds for Inhibiting Soluble Epoxide Hydrolases:

In addition to the methods provided above, the present inventionprovides in another aspect, compounds that can inhibit the activity ofsoluble epoxide hydrolases. In particular, the present inventionprovides compounds having a formula selected from formula (I) above.

In one embodiment, compounds are those compounds described above as forthe recited uses.

In one embodiment, sEH inhibitors for treating hypertension or highblood pressure have an IC₅₀ in a defined assay of less than 50 μM. Inanother embodiment, the compounds have an IC₅₀ of 1 μM or less. Inanother embodiment, the compounds have an IC₅₀ of 500 nM or less. Inanother embodiment, the compounds have an IC₅₀ of 150 nM or less. Inanother embodiment, the compounds have an IC₅₀ of 100 nM or less. Inanother embodiment, the compounds have an IC₅₀ of 50 nM or less. Inanother embodiment, the compounds have an IC₅₀ of 1 nM or less.

Methods of Preparation

The compounds of the present invention can be prepared by a variety ofmethods as outlined generally in the scheme below. It should be notedthat the synthetic conditions illustrated in the following scheme arealso applicable to those inhibitors based on 4-aminomethylpiperidine(those with a CH₂ spacer).

Scheme 1—Introduction of a Heterocyclic Pharmacophore

Scheme 1 illustrates general methods that can be used for preparation ofcompounds of the invention having heterocyclic secondary pharmacophore,for example a piperidine. While the scheme is provided for the synthesisof N-(1-benzoylpiperidin-4-yl)-N′-(adamant-1-yl)ureas, one of skill inthe art will understand that a number of commercially available orsynthetic heterocyclic amines could be used in place of4-aminopiperidine, and that other substituents other than benzoyl couldalso be employed.

As shown in Scheme 1, 4-aminopiperidine (available from Aldrich ChemicalCo., Milwaukee, Wis., USA) is combined with benzaldehyde at roomtemperature to provide intermediate (i). BOC protection of thepiperidine nitrogen provides intermediate carbamate (ii). Reaction of(ii) with a suitable isocyanate provides intermediate (iii).Deprotection of the piperidine (iv) and reaction with a suitablealkylating or acylating agent provides the target compounds.Substitution of adamantyl isocyanate with, for example, a substituted orunsubstituted phenyl isocyanate or cycloalkyl isocyanate (e.g.cyclohexyl isocyanate, also available from Aldrich Chemical Co.)provides other compounds of the invention.

The following examples are provided to illustrate the invention and arenot intended to limit any aspect of the invention as set forth above orin the claims below.

EXAMPLES

All melting points were determined with a Thomas-Hoover apparatus (A.H.Thomas Co.) and are uncorrected. Compounds with no melting point valuesexist in the solid state as either foams or glassy solids. Mass spectrawere measured by LC-MS (Waters 2790). ¹H-NMR spectra were recorded onQE-300 spectrometer, using tetramethylsilane as an internal standard.Signal multiplicities are represented as singlet (s), doublet (d),double doublet (dd), triplet (t), quartet (q), quintet (quint),multiplet (m), broad (br), broad singlet (brs), broad doublet (br d),broad triplet (br t), broad multiplet (br m), doublet of doublet ofdoublets (ddd) and quartet of doublets (qd). Synthetic methods aredescribed for representative compounds.

The abbreviations used in the examples below have the following meaning:melting point (Mp), mass spectroscopy (MS), thin layer chromatography(TLC), the parent peak in the MS plus H⁺ ([M+H]⁺), minute (min),kilogram (kg), milligram (mg), nanomolar (nM), tetrahydrofuran (THF),tertiary butoxy carbonyl (BOC), potassium sulfate (KHSO₄), potassiumhydroxide (KOH), magnesium sulfate (MgSO₄), hydrogen chloride (HCl),dimethylsulfoxide (DMSO), ethyl (Et), ethyl acetate (EtOAc), methanol(MeOH), dichloromethane (CH₂Cl₂, DCM), area under the concentration(AUC).

Lower case bolded Roman numerals in the examples below refer to thecorresponding intermediates in Scheme 1 above. Compounds numbers arealso used as provided in the Schemes as well as in the Tables below.

Example 1

4-Aminopiperidine (2.125 g, 21.2 mmol) was dissolved in toluene (50 mL).To this was added benzaldehyde (2.16 mL, 21.2 mmol). The reaction fittedwith a Dean-Stark trap and a condenser and was refluxed for 4 hoursunder an atmosphere of nitrogen. At this point, when no additional waterwas seen to form, the reaction was cooled to 0° C. and BOC anhydride(4.63 g, 21.2 mmol) was added via syringe over 10 minutes. The reactionwas allowed to warm to room temperature over 1 hr and was stirred for anadditional 12 hrs. The solvent was removed in vacuo and the resultingoil was treated with KHSO₄(aq) (1 M, 21.2 mL). This was stirred for 1.5hours. Water (25 mL) was added to the reaction and the aqueoussuspension washed with diethylether (3×100 mL). The water layer was thenbasified to pH=10 with KOH (s) and was extracted with dichloromethane(3×100 mL). The organic layer was dried over MgSO₄ and evaporated togive 4.76 g of a yellow oil. To this oil (1.0 g) was added THF (25 mL).This was stirred for 5 minutes until the oil was completely dissolved.1-Adamantylisocyanate (0.886 mg, 5.0 mmol, 1 eq) was added and thereaction stirred overnight under an atmosphere of nitrogen. The solventwas removed and the residue was chromatographed on silica with 1:1ethylacetate:hexanes. The major fraction was collected (TLC rf=0.8 1:1hexane:EtOAc) and the solvent removed. The resultant residue was treatedwith a solution of HCl in methanol (35 mL, 4M). This was stirred for 12hours. The solvent was removed to give the product, after drying at 80°C. under vacuum, as a white powder (1.123 g, 73% yield overall).

N-(piperidin-4-yl)-N′-(adamant-1-yl)urea hydrochloride (iv, 1175)

¹H (300 MHz, DMSO d6): 8.96 (br 2H), 6.22 (br, 6H, urea NH+H₂O),3.61-3.52 (m, 1H), 3.24-3.10 (m, 2H), 2.95-2.80 (m, 2H), 2.10-1.70 (brm, 11H), 1.70-1.40 (br m, 8H).

N-((piperidin-4-yl)methyl)-N′-(adamant-1-yl)urea hydrochloride (1118)

This was run as per above with a yield of 95%. Mp. (free base): 199-201°C. dec.

¹H NMR (300 MHz, DMSO): 8.79 (br, 1H), 8.50 (br, 1H), 6.00 (br, 1H),5.80 (br, 1H), 3.20 (br d, J=12.3 Hz, 2H), 2.80-2.70 (br m, 3H),2.00-1.40 (br m, 19H), 1.30-1.15 (br m, 2H).

Example 2 General Procedure for the Alkylation of Piperidinyl Ureas:N-(1-ethylpiperidin-4-yl)-N′-(adamant-1-yl)urea (R=Et, 1152)

The appropriate piperidinyl urea (0.319 mmol) was combined with theappropriate alkyl or benzyl bromide (X=Br) (0.382 mmol) and K₂CO₃ (132mg, 0.96 mmol) in DMF (3.0 mL). The reaction was heated at 50° C. for 12hours. At this point, the reaction was cooled to room temperature andthe solvent was removed in vacuo. The residue was partitioned betweenDCM and aqueous NaHCO₃ (satd) and the organic layer removed and driedwith Na₂SO₄. The solvent was evaporated and the residue chromatographedon silica gel using ammonia saturated methanol/DCM as the eluent(5:100). Yield=42%. Mp.: 203-213° C. dec. ¹H NMR (300 MHz, CDCl₃):4.15-4.05 (br, 2H), 3.63-3.47 (m, 1H), 2.91-2.81 (br m, 2H), 2.39 (q,J=7.18 Hz, 2H), 2.13-1.88 (br m, 13H), 1.66 (br, 6H), 1.40 (qd, J=8.3,3.3 Hz, 2H), 1.07 (t, J=7.19 Hz, 3H).

Example 3

N-(1-n-propylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1155)

Yield=60%. Mp.: 195-200° C. dec. ¹H (300 MHz, CDCl₃): 4.10-4.00 (br,2H), 3.60-3.45 (m, 1H), 2.90-2.78 (m, 2H), 2.32-2.22 (m, 2H), 2.10-1.70(m, 13H), 1.70-1.57 (br, 6H), 1.56-1.30 (m, 4H), 0.88 (t, J=7.4 Hz, 3H).

Example 4

N-(1-n-butylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1160)

Yield=53%. Mp.: 195-200° C. dec. ¹H (300 MHz, CDCl₃): 4.05-3.95 (br,2H), 3.51-3.45 (m, 1H), 2.90-2.80 (m, 2H), 2.35-2.25 (m, 2H), 2.10-1.60(br m, 19H), 1.50-1.25 (m, 6H), 0.89 (t, J=7.2 Hz, 3H).

Example 5

N-(1-benzylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1158)

Yield=46%. Mp.: 170-173° C. ¹H (300 MHz, CDCl₃): 7.35-7.20 (m, 5H),4.00-3.94 (br, 2H), 3.58-3.45 (m, 1H), 3.43 (s, 2H), 2.80-2.72 (m, 2H),2.10-1.60 (br m, 19H), 1.35 (qd, J=7.9, 3.3 Hz, 2H).

Example 6

N-((1-ethylpiperidin-4-y)methyl)-N′-(adamant-1-yl)urea (1154)

Yield=50%. Mp.: 143-151° C. dec. ¹H (300 MHz, CDCl₃): 4.28 (t, J=5.4 Hz,1H), 4.09 (br, 1H), 3.05 (t, J=6.2 Hz, 2H), 2.98-2.89 (br m, 2H), 2.38(q, J=7.4 Hz, 2H), 2.10-1.60 (br m, 19H), 1.52-1.40 (br m, 1H), 1.27(qd, J=12.4, 3.7 Hz, 2H), 1.08 (t, J=7.2 Hz, 3H).

Example 7

N-((1-n-propylpiperidin-4-y)methyl)-N′-(adamant-1-yl)urea (1122)

Yield=40%. ¹H (300 MHZ, CDCl₃): 4.69 (t, J=5.8 Hz, 1H), 4.38 (br, 1H),3.08-2.94 (m, 4H), 2.42-2.32 (m, 2H), 2.10-1.55 (br m, 22H), 1.36 (qd,J=11.8, 3.3 Hz, 2H), 0.89 (t, J=7.4 Hz, 3H).

Example 8

N-((1-n-butylpiperidin-4-yl)methyl)-N′-(adamant-1-yl)urea (1161)

Yield=43%. ¹H (300 MHz, CDCl₃): 4.30 (br, 1H), 4.12 (br, 1H), 3.05 (t,J=6.2 Hz, 2H), 2.98-2.88 (m, 2H), 2.34-2.26 (m, 2H), 2.10-1.2 (br m,26H), 0.09 (t, J=7.2 Hz, 3H).

Example 9

N-((1-benzylpiperidin-4-yl)methyl)-N′-(adamant-1-yl)urea (1119)

Yield=48%. Mp.: 162-167° C. ¹H (300 MHz, CDCl₃): 7.35-7.20 (m, 5H), 4.37(br t, J=5.8 Hz, 1H), 4.17 (br, 1H), 3.48 (s, 2H), 2.99 (t, J=6.2 Hz,2H), 2.95-2.80 (br m, 2H), 2.10-1.40 (br m, 20H), 1.27 (qd, J=11.9, 3.5Hz, 2H).

Example 10A

General Procedure for the Acylation of Piperidines:

N-(1-acetylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1153)

The desired piperidinyl urea (6.6 mmol) and an appropriate carboxylicacid (or ester-acid) (7.92 mmol), DMAP (0.805 g, 6.6 mmol) and TEA (5.0mL, 36 mmol) were all combined in dichloromethane at 0° C. The reactionwas allowed to stir for 10 minutes. At this point, EDCI (1.38 g, 7.26mmol) was added and the reaction was allowed to warm to rt over 2 hours.After reaching room temperature, the reaction was allowed to stir for 18hrs. The reaction was then washed with K₂CO₃(aq) (1M, 3×50 mL) followedby HCl_((aq)) (1M, 3×50 mL). The organic layer was dried and evaporatedto give a yellow oil. Recrystallization from acetone or chromatography(SiO₂) with 5% MeOH/DCM afforded the product. Yield=75%. Mp.: 205-206°C. ¹H (300 MHz, CDCl₃): 4.67 (br d, J=6.9 Hz, 1H), 4.57 (br s, 1H), 4.44(br d, J=13.1 Hz, 1H), 3.90-3.65 (m, 2H), 3.13 (br t, J=13.1 Hz, 1H),2.74 (br t, J=13.2 Hz, 1H), 2.20-1.50 (br m, 20H), 1.30-1.10 (m, 2H).

Example 10B Alternative Synthesis ofN-(1-Acetylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1153) Preparation ofN-Acetyl piperid-4-yl amide

A reactor was charged with 1.00 mole-equivalent of4-piperidinecarboxamide, 15.9 mole-equivalents of THF, and 1.23mole-equivalents of N,N-(diisopropyl)ethylamine under a nitrogenatmosphere. The resulting mixture was cooled to 20° C. internal, and1.10 mole-equivalents of acetic anhydride was added at such a rate as tomaintain an internal temperature of less than 30° C. After addition wascomplete, the reaction mixture was stirred while maintaining an internaltemperature of 20° C. The reaction contents was monitored until theamount of unreacted 4-piperidinecarboxamide was less than 1% relative toN-acetyl piperid-4-yl amide product (typically about 4-10 hours). Theprecipitated product was collected by filtration and washed with THF toremove excess (diisopropyl)ethylamine hydrochloride. The solid productwas dried to constant weight in a vacuum oven under a nitrogen bleedwhile maintaining an internal temperature of ≦50° C. to afford theproduct as a white solid in 94% yield.; Mp.: 172-174° C. ¹H NMR(CD₃OD)δ: 4.48-4.58 (bd, 1H), 3.92-4.01 (bd, 1H), 3.08-3.22 (m, 1H), 2.62-2.74(m, 1H), 2.44-2.53 (m, 1H), 2.12 (s, 3H), 1.88-1.93 (m, 2H), 1.45-1.72(m, 2H); MS: 171 [M+H]⁺.

Preparation of N-(1-Acetylpiperidin-4-yl)-N′-(adamant-1-yl)urea

A reactor was charged with 1.00 mole-equivalents of N-acetylpiperid-4-yl amide, 0.87 mole-equivalents of 1-adamantyl amine, and 49.7mole-equivalents of acetonitrile, and the resulting mixture was heatedto 75° C. internal under a nitrogen atmosphere. (Diacetoxyiodo)benzene(1.00 mole-equivalents) was charged portionwise in such a way that thereaction mixture was maintained between 75-80° C. internal. After the(diacetoxyiodo)benzene was added, the reaction mixture was heated to 80°C. internal. The reaction contents was monitored until the amount ofunreacted 1-adamantyl amine was less than 5% relative to productN-(1-acetylpiperidin-4-yl)-N′-(adamant-1-yl)urea (typically about 1-6hours). After completion, the reaction mixture was cooled to 25° C.internal, and approximately 24 mole-equivalents of solvent was distilledout under vacuum while maintaining internal temperature below 40° C. Thereaction mixture was cooled with agitation to 0-5° C. internal andstirred for an additional 2 hours. The technical product was collectedby filtration and washed with acetonitrile. The crude product was driedto constant weight in a vacuum oven under a nitrogen bleed maintainingan internal temperature of ≦50° C. The dried, crude product was slurriedwith water maintaining an internal temperature of 20±5° C. internal for4 hours and then collected by filtration. The filter cake washed withheptane under a nitrogen atmosphere then dried to constant weight in avacuum oven under a nitrogen bleed maintaining an internal temperatureof ≦70° C. to afford product as a white solid in 72% yield based on1-adamantyl amine. ¹H NMR (DMSO-d₆) δ: 5.65-5.70 (bd, 1H), 5.41 (s, 1H),4.02-4.10 (m, 1H), 3.61-3.70, (m, 1H), 3.46-3.58 (m, 1H), 3.04-3.23 (m,1H), 2.70-2.78 (m, 1H), 1.98 (s, 3H), 1.84 (s, 6H), 1.64-1.82 (m, 2H),1.59 (s, 6H), 1.13-1.25 (m, 1H), 1.00-1.12 (m, 1H); MS: 320 [M+H]⁺; m.p.202-204° C.

Example 11

N-(1-propionylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1163)

Made by treating 1 eq of the piperidine with 1 eq of propanoyl chloridein pyridine ([starting material]=0.10 M) at 0° C. for 12 hrs. Afterremoval of the solvent, the product was chromatographed on silica gelwith 90:1 DCM: MeOH/NH₃ to give to target in 20% yield. Mp.: 211-224° C.dec. ¹H (300 MHz, CDCl₃): 4.52 (br d, J=12.6 Hz, 1H), 4.40-4.00 (br,2H), 3.90-3.70 (m, 2H), 3.10 (br t, J=12.4 Hz, 1H), 2.75 (br t, J=12.5Hz, 1H), 2.34 (q, J=7.4 Hz, 2H), 2.10-1.60 (br m, 17H), 1.30-1.15 (m,2H), 1.13 (t, J=7.3 Hz, 3H).

Example 12

N-(1-butyrylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1157)

Synthesized as per 1163. Yield: 71%. Mp.: 148-188° C. dec. ¹H NMR (300MHz, CDCl₃): 4.52 (br d, J=13.3, 11H), 4.25-4.10 (br, 2H), 3.85-3.65(br, 2H), 3.10 (br t, J=11.5 Hz, 1H), 2.75 (br t, J=11.3 Hz, 1H),2.35-2.23 (m, 2H), 2.10-1.60 (br m, 19H), 1.30-1.15 (m, 2H), 0.96 (t,J=7.4 Hz, 3H).

Example 13

N-(1-benzoylpiperidin-4-yl)-N′-(adamant-1-yl)urea (1159)

Yield=63% (via acyl chloride). ¹H (300 MHz, CDCl₃): 7.44-7.32 (m, 5H),5.00-4.50 (br m, 3H), 3.90-3.78 (br, 1H), 3.76-3.60 (br, 1H), 3.20-2.90(br, 2H), 2.10-1.60 (br m, 17H), 1.50-1.20 (br m, 2H).

Example 14

N-(1-(Pyridine-2-carbonyl)piperidin-4-yl)-N′-(adamant-1-yl)urea (1201)

Yield=70% via EDCI coupling (see 1153). ¹H (300 MHz, CDCl₃): 8.59 (br d,J=5.0 Hz, 1H), 7.80 (td, J=7.7, 1.7 Hz, 1H), 7.56 (br d, J=7.6 Hz, 1H),7.35 (ddd, J=7.6, 4.8, 1.2 Hz, 1H), 4.70-4.50 (br m, 3H), 3.90-3.70 (m,2H), 3.15 (br t, J=12.5 Hz, 1H), 2.95 (br t, J=12.3 Hz, 1H), 2.10-1.60(br m, 17H), 1.50-1.20 (br m, 2H).

N-(1-(Pyridine-3-carbonyl)piperidin-4-yl)-N′-(adamant-1-yl)urea (1434)

Yield=86% via EDCI coupling. ¹H (300 MHz, CDCl₃): 8.67 (br d, J=5.0 Hz,1H), 8.65 (br, 1H), 7.74 (br d, J=8.0 Hz, 1H), 7.38 (dd, J=7.9, 5.0 Hz,1H), 4.67-4.23 (br m, 3H), 3.94-3.70 (m, 1H), 3.70-3.55 (br, 1H),3.20-2.90 (br m, 2H), 2.10-1.60 (br m, 17H), 1.50-1.20 (br m, 2H).

Example 15

N-(1-(pyridine-4-carbonyl)piperidin-4-yl)-N′-(adamant-1-yl)urea (1433)

Yield=81% via EDCI coupling. Mp 197-199° C. ¹H (300 MHz, CDCl₃): 8.70(m, 2H), 7.26 (m, 2H), 4.60 (br d, J=14.2 Hz, 1H), 4.40 (d, J=7.6 Hz,1H), 4.31 (s, 1H), 3.90-3.70 (m, 1H), 3.57 (br d, J=14.0 Hz, 1H), 3.13(br t, J=12.3 Hz, 1H), 2.95 (br t, J=12.0 Hz, 1H), 2.10-1.60 (br m,17H), 1.37 (m, 1H), 1.21 (m, 1H).

Example 16

N-((1-acetylpiperidin-4-yl)methyl)-N′-(adamant-1-yl)urea (1156)

Yield=55%. ¹H (300 MHz, CDCl₃) 5.10-4.50 (br, 2H), 4.60 (d, J=13.3 Hz,1H), 3.81 (d, J=13.4 Hz, 1H), 3.15 (br dd, J=13.7, 4.4 Hz, 1H), 3.03 (brt, J=12.6 Hz, 1H), 2.92 (br dd, J=13.0, 4.5 Hz, 1H), 2.53 (br t, J=12.9Hz, 1H), 2.4-1.4 (br m, 21H), 1.20-0.99(m, 2H).

Example 17

N-((1-propanoylpiperidin-4-yl)methyl)-N′-(adamant-1-yl)urea (1162)

Yield=20% (via acid chloride). ¹H (300 MHz, CDCl₃): 4.60 (br d, J=12.0Hz, 1H), 3.85 (br d, J=12.3 Hz, 1H), 3.20-2.80 (br, 3H), 2.52 (br t,J=12.8 Hz, 1H), 2.33 (q, J=7.5 Hz, 2H), 2.4-1.4 (br m, 18H), 1.13 (t,J=7.5 Hz, 3H), 1.15-1.05 (br m, 2H). (note, sample contained water,therefore no urea N—H are seen).

Example 18

N-((1-butyrylpiperidin-4-yl)methyl)-N′-(adamant-1-yl)urea (1120)

Yield=35%. Mp.: 117-149° C. dec. ¹H (300 MHz, CDCl₃): 4.78 (br t, J=4.7Hz, 1H), 4.61 (br d, J=13.1 Hz, 1H), 4.47 (s, 1H), 3.86 (br d, J=13.6Hz, 1H), 3.20-3.08 (m, 1H), 2.98 (t, J=13.1 Hz, 1H), 2.95-2.84 (m, 1H),2.52 (t, J=12.6 Hz, 1H), 2.29 (t, J=7.4 Hz, 2H), 2.10-1.50 (m, 20H),1.20-1.00 (m, 2H), 0.96 (t, J=7.4 Hz, 3H).

Example 19

N-((1-benzoylpiperidin-4-yl)methyl)-N′-(adamant-1-yl)urea (1121)

Yield=45%. ¹H (300 MHz, CDCl₃): 7.45-7.34 (m, 5H), 4.80-4.60 (br, 1H),4.50-4.40 (br, 1H), 4.30-4.05 (br, 1H), 3.80-3.60 (br, 1H), 3.20-2.60(br, 4H), 2.10-1.5 (br m, 18H), 1.30-1.0 (br, 2H).

Example 20

N-((1-(pyridine-2-carbonyl)piperidin-4-yl)methyl)-N′-(adamant-1-yl)urea(1207)

Yield=73%. ¹H (300 MHz, CDCl₃): 8.59 (br d, J=5.0 Hz, 1H), 7.79 (td,J=7.7, 1.7 Hz, 1H), 7.56(br d, J=7.7 Hz, 1H), 7.33 (ddd, J=7.6, 4.8, 1.2Hz, 1H), 4.70(br d, J=12.7 Hz, 1H), 4.47 (br m, 1H), 4.20 (s, 1H), 3.88(br d, 13.1 Hz, 1H), 3.20-2.90 (m, 3H), 2.77 (br t, J=12.6 Hz, 1H),2.10-1.50 (m, 18H), 1.15-1.05 (m, 2H).

Example 21

N-((1-(pyridine-3-carbonyl)piperidin-4-yl)methyl)-N′-(adamant-1-yl)urea(1436)

Yield=quantitative. ¹H (300 MHz, CDCl₃): 8.65 (dd, J=4.9, 1.6 Hz, 1H),8.64 (d, J=2.0 Hz, 1H), 7.74 (dt, 7.8, 1.9 Hz, 1H), 7.37 (dd, J=7.9, 4.9Hz, 1H), 5.00-4.90 (br, 1H), 4.78-4.60 (br, 1H), 4.60-4.44 (br 1H),3.79-3.62 (br, 1H), 3.21-2.68 (br m, 4H), 2.10-1.50 (m, 18H), 1.15-1.05(m, 2H).

Example 22

N-((1-(pyridine-4-carbonyl)piperidin-4-yl)methyl)-N′-(adamant-1-yl)urea(1435)

Yield=77%. 8.70-8.66 (m, 2H), 7.28-7.25 (m, 2H), 4.77-4.58 (br, 2H),4.44-4.36 (br, 1H), 3.60 (br d, J=13.5 Hz, 1H), 3.20-2.95 (m, 3H), 2.77(br t, J=12.5 Hz, 1H), 2.10-1.50 (m, 18H), 1.15-1.05 (m, 2H).

Example 23

4-[4-(3-Adamantan-1-yl-ureido)-piperidin-1-yl]-4-oxo-butanoic acidmethyl ester (1205)

Yield=78%. Mp 169-175° C. dec. ¹H (300 MHz, CDCl₃): 4.65-4.34 (br m,3H), 3.90-3.67 (br m, 2H), 3.69 (s, 3H), 3.12 (br t, J=13.2 Hz, 1H),2.76 (br t, J=13.2 Hz, 1H), 2.71-2.54 (m, 4H), 2.20-1.5 (m, 17H),1.30-1.10 (m, 2H).

Example 24

5-[4-(3-Adamantan-1-yl-ureido)-piperidin-1-yl]-5-oxo-pentanoic acidmethyl ester (1206)

Yield=61%. Mp 152-154° C. ¹H (300 MHz, CDCl₃): 4.65-4.34 (br m, 3H),3.90-3.67 (br m, 2H), 3.66 (s, 3H), 3.09 (br t, J=13.7 Hz, 1H), 2.70 (brt, J=13.7 Hz, 1H), 2.45-2.31 (m, 4H), 2.20-1.5 (m, 19H), 1.30-1.10 (m,2H).

Example 25

2-[4-(3-Adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acidmethyl ester (1202)

Yield=63%. ¹H (300 MHz, CDCl₃):8.03 (d, J=7.9 Hz, 1H), 7.58 (t, J=7.7Hz, 1H), 7.46 (t, J=7.7 Hz, 1H), 7.25 (d, J=7.7 Hz, 1H), 5.00-4.62 (br,2H), 4.55 (br d, J=13.0 Hz, 1H), 3.87 (s, 3H), 3.85-3.72 (br m, 1H),3.13 (br d, J=13.1 Hz, 1H), 3.11-2.94 (m, 2H), 2.10-1.10 (m, 19H).

Example 26

3-[4-(3-Adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acidmethyl ester (1203)

Yield=61%. ¹H (300 MHz, CDCl₃): 8.10 (dd, J=7.6, 1.4 Hz, 1H), 8.04 (d,J=1.4 Hz, 1H), 7.58 (dd, J=7.6, 1.4 Hz, 1H), 7.50 (t, J=7.6 Ha, 1H),4.7-4.4 (br, 3H), 3.93 (s, 3H), 3.90-3.81 (br, 1H), 3.70-3.55 (br, 1H),3.20-3.90 (br m, 2H), 2.15-1.60 (br m, 17H), 1.50-1.10 (br m, 2H).

Example 27

4-[4-(3-Adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acidmethyl ester (1204)

Yield=70%. Mp 239-243° C. ¹H (300 MHz, CDCl₃): 8.08 (d, J=8.5 Hz, 2H),7.43 (d, J=8.5 Hz, 2H), 4.67-4.50 (br m, 2H), 4.45 (br, 1H), 3.94 (s,3H), 3.90-8.74 (m, 1H), 3.65-3.55 (br m, 1H),), 3.20-3.90 (br m, 2H),2.15-1.60 (br m, 17H), 1.50-1.10 (br m, 2H).

Example 28

4-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidin-1-yl}-4-oxo-butanoicacid methyl ester (1208)

Yield=72%. ¹H (300 MHz, CDCl₃): 4.70-4.10 (br, 2H), 4.58 (d, J=12.4 Hz,1H), 3.89 (d, J=12.5 Hz, 1H), 3.69 (s, 3H), 3.15 (br dd, J=13.7, 4.4 Hz,1H), 3.03 (br t, J=12.6 Hz, 1H), 2.92 (br dd, J=13.0, 4.5 Hz, 1H), ),2.64 (s, 4H), 2.53 (br t, J=12.9 Hz, 1H), 2.10-1.50 (br m, 18H),1.15-1.00 (m, 2H).

Example 29

5-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidin-1-yl}-5-oxo-pentanoicacid methyl ester (1212)

Yield=42%. ¹H (300 MHz, CDCl₃): 4.70-4.10 (br m, 2H), 4.58 (d, J=12.4Hz, 1H), 3.89 (d, J=12.5 Hz, 1H), 3.66 (s, 3H), 3.15 (br dd, J=13.7, 4.4Hz, 1H), 3.03 (br t, J=12.6 Hz, 1H), 2.92 (br dd, J=13.0, 4.5 Hz, 1H),), 2.53 (br t, J=12.9 Hz, 1H), 2.39 (m, 4H), 2.10-1.50 (br m, 20H),1.20-0.95 (m, 2H).

Example 30

2-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidine-1-carbonyl}-benzoicacid methyl ester (1210)

Yield=76%. ¹H (300 MHz, CDCl₃): 8.02 (d, J=7.8 Hz, 1H), 7.57 (td, J=7.5,1.2 Hz, 1H), 7.45 (td, J=7.6, 1.2 Hz, 1H), 7.26 (d, J=7.6 Hz, 1H),5.10-4.85 (br m, 1H), 4.74 (br d, J=12.5 Hz, 1H), 4.70-4.60 (br, 1H),3.87 (s, 3H), 3.35 (br d, J=12.5 Hz, 1H), 3.20-3.10 (m, 1H), 3.00-2.70(m, 3H),), 2.10-1.50 (br m, 18H), 1.20-0.95 (m, 2H).

Example 31

3-(4-[(3-Adamantan-1-yl-ureido)-methyl-piperidine-1-carbonyl]-benzoicacid methyl ester (1209)

Yield=67%. ¹H (300 MHz, CDCl₃): 8.08 (d, J=7.7 Hz, 1H), 8.04 (s, 1H),7.58 (d, J=7.7 Hz, 1H), 7.49 (t, J=7.6 Hz, 1H), 5.10-4.40 (br m, 3H),3.93 (s, 3H) 3.75-3.63 (br, 1H), 3.20-2.80 (br m, 4H),), 2.10-1.50 (brm, 18H), 1.20-0.95 (m, 2H).

Example 32

4-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidine-1-carbonyl}-benzoicacid methyl ester (1211)

Yield=71%. ¹H NMR (300 MHz, CDCl₃): 8.07 (d, J=8.5, 2H), 7.44 (d, J=8.5,2H), 4.70 (br d, J=12.1 Hz, 1H), 4.55-4.45 (br, 1H), 4.22 (br, 1H), 3.94(s, 3H), 3.64 (br d, J=12.4 Hz, 1H), 3.25-2.70 (br m, 4H), 2.10-1.50 (brm, 18H), 1.20-0.95 (m, 2H).

General Procedure for the Hydrolsis of Methyl Esters to theCorresponding Acids.

The parent ester was dissolved in methyl alcohol to a concentration of 1M. To this was added 1.2 eq of KOH (as a 4 M solution). The reaction washeated to 60° C. for 6 hrs. The solvent was removed and the residuechormatographed on silica gel using a 94:5:1 DCM:MeOH:HOAc eluent.Yields were greater than 90%.

Example 33

4-[4-(3-Adamantan-1-yl-ureido)-piperidin-1-yl]-4-oxo-butanoic acid(1503)

Mp 196° C. dec. ¹H NMR (300 MHz, DMSO): 12.14 (br, 1H), 5.67 (d, J=7.8Hz, 1H), 5.40 (s, 1H), 4.05 (br d, J=13.2 Hz, 1H), 3.71 (br d, J=13.5Hz, 1H), 3.50 (br, 1H), 3.07 (br t, J=10.8 Hz, 1H), 2.74 (br t, J=10.8Hz, 1H), 2.50-2.30 (m, 4H), 2.00-1.60 (br m, 17H), 1.40-0.90 (m, 2H).

Example 34

5-[4-(3-Adamantan-1-yl-ureido)-piperidin-1-yl]-5-oxo-pentanoic acid(1501)

¹H NMR (300 MHz, DMSO): 12.11 (br, 1H), 5.66 (br d, J=7.1 Hz, 1H), 5.40(br s, 1H), 4.08 (br d, J=13.1 Hz, 1H), 3.67 (br d, J=13.6 Hz, 1H),3.58-3.41 (br, 1H), 3.05 (br t, J=11.7 Hz, 1H), 2.73 (br t, J=11.7 Hz,1H), 2.28 (t, J=7.4 Hz, 2H), 2.21 (t, J=7.4 Hz, 2H), 2.00-1.50 (br m,19H), 1.20-0.90 (m, 2H).

Example 35

2-[4-(3-Adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acid(1507)

Mp 219° C. dec. ¹H NMR (300 MHz, DMSO): 13.16 (br, 1H), 7.90 (dd, J=7.7,0.89 Hz, 1H), 7.62 (td, J=7.5, 0.9 Hz, 1H), 7.49 (td, J=7.6, 1.0 Hz,1H), 7.27 (d, J=7.3 Hz, 1H), 5.71 (d, J=7.4 Hz, 1H), 5.45 (s, 1H),4.24-4.12 (br m, 1H), 3.60-3.45 (br, 1H), 3.22-3.10 (br m, 1H),3.05-2.85 (br, 2H), 2.02-1.38 (br m, 17H), 1.35-1.06 (br m, 2H).

Example 36

3-[4-(3-Adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acid(1505)

¹H NMR (300 MHz, DMSO): 13.23 (br, 1H), 7.97 (br d, J=7.4 Hz, 1H), 7.86(br s, 1H), 7.59 (br d, J=7.3 Hz, 1H), 7.54 (t, J=7.3 Hz, 1H), 5.70 (brd, 7.8 Hz, 1H), 5.42 (br s, 1H), 4.30-4.10 (br, 1H), 3.65-2.95 (br m,4H), 2.00-1.50 (br m, 17H), 1.35-1.06 (br m, 2H).

4-[4-(3-Adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acid(1523)

Mp 245-251° C. ¹H NMR (300 MHz, DMSO): 13.12 (br, 1H), 7.95 (d, J=7.5Hz, 1H), 7.45 (d, J=7.5 Hz, 1H), 5.69 (d, J=5.7 Hz, 1H), 5.44 (s, 1H),4.29-4.11 (br, 1H), 3.68-2.88 (br m, 4H), 2.00-1.50 (br m, 17H),1.35-1.06 (br m, 2H).

Example 37

4-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidin-1-yl}-4-oxo-butanoicacid (1502)

¹H NMR (300 MHz, DMSO): 12.70-10.93 (br, 1H), 5.71 (br t, J=5.2 Hz, 1H),5.44 (s, 1H), 4.30 (br d, J=11.4 Hz, 1H), 3.83 (br d, J=12.0 Hz, 1H),2.91 (br t, J=12.9 Hz, 1H), 2.84-2.78 (m, 2H), 2.50-2.30 (br m, 5H),2.10-1.50 (br m, 18H), 1.15-0.80 (m, 2H).

Example 38

5-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidin-1-yl}-5-oxo-pentanoicacid (1500)

¹H NMR (300 MHz, DMSO): 12.80-11.10 (br, 1H), 5.70 (br t, J=5.6 Hz, 1H),5.44 (s, 1H), 4.33 (br d, J=12.4 Hz, 1H), 3.80 (br d, J=13.0 Hz, 1H),2.89 (br t, J=12.8 Hz, 1H), 2.83-2.76 (m, 2H), 2.44 (br t, J=12.6 Hz,1H), 2.27 (t, J=7.5 Hz, 2H), 2.21 (t, J=7.4 Hz, 2H), 2.10-1.50 (br m,20H), 1.20-0.95 (m, 2H).

Example 39

2-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidine-1-carbonyl}-benzoicacid (1506)

Mp 192° C. dec. ¹H NMR (300 MHz, DMSO): 13.60-11.60 (br, 1H), 7.87 (brd, J=7.7 Hz, 1H), 7.53 (br t, J=7.4 Hz, 1H), 7.42 (br t, J=7.4 Hz, 1H),7.18 (br d, J=7.4 Hz, 1H), 5.75 (br m, 1H), 5.47 (br, 1H), 4.45 (br d,J=12.4 Hz, 1H), 3.17 (br d, J=11.5 Hz, 1H), 2.90-2.55 (br m, 4H),2.10-1.50 (br m, 18H), 1.20-0.95 (m, 2H).

Example 40

3-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidine-1-carbonyl}-benzoicacid (1504)

¹H NMR (300 MHz, DMSO): 13.53-12.70 (br, 1H), 7.97 (d, J=7.3 Hz, 1H),7.85 (s, 1H), 7.61-7.51 (m, 2H), 5.75-5.68 (br m, 1H), 5.43 (s, 1H),4.50-4.37 (br, 1H), 3.50-2.55 (br m, 5H), 2.10-1.50 (br m, 18H),1.20-0.92 (m, 2H).

Example 41

4-{4-[(3-Adamantan-1-yl-ureido)-methyl]-piperidine-1-carbonyl}-benzoicacid (1522)

Mp 147° C. dec. ¹H NMR (300 MHz, DMSO): 13.80-12.40 (br, 1H), 7.96 (d,J=8.1 Hz, 1H), 7.43 (d, J=8.2 Hz, 1H), 5.72 (t, J=5.8 Hz, 1H), 5.45 (s,1H), 4.44 (br d, J=11.5 Hz, 1H), 3.50 (br m, 1H), 3.50-2.55 (br m, 5H),2.10-1.50 (br m, 18H), 1.20-0.90 (m, 2H).

Example 42

N-(1-Methanesulfonyl piperidin-4-yl)-N′-(adamant-1-yl)ureaN-Methanesulfonyl piperid-4-yl amide

A reactor was charged with 1.0 mole-equivalent of4-piperidinecarboxamide, 16.4 mole-equivalents of THF, and 1.2mole-equivalents of N,N-(diisopropyl)ethylamine under a nitrogenatmosphere. The resulting mixture was cooled to 0-5° C. internal, and1.2 mole-equivalents of methanesulfonyl chloride was added at such arate as to maintain an internal temperature of less than 10° C. Afteraddition was complete, the reaction mixture was stirred allowing thetemperature to rise to 20° C. internal. The reaction contents wasmonitored until the amount of unreacted 4-piperidinecarboxamide was lessthan 1% relative to N-methanesulfonyl piperid-4-yl amide product(typically about 2-12 hours). The precipitated product was collected byfiltration then washed with dichloromethane to remove excess(diisopropyl)ethylamine hydrochloride. The solid product was dried toconstant weight in a vacuum oven under a nitrogen bleed maintaining aninternal temperature of ≦50° C. to afford product as a light yellowsolid in 87% yield. Mp.: 126-128° C. ¹H NMR (DMSO-d₆) δ: 7.30 (s, 1H),6.91 (s, 1H), 3.46-3.59 (m, 2H), 2.83 (s, 3H), 2.60-2.76 (m, 2H),2.08-2.24 (m, 1H), 1.70-1.86 (m, 2H), 1.43-1.62 (m, 2H); MS: 207 [M+H]⁺.

N-(1-Methanesulfonyl piperidin-4-yl)-N′-(adamant-1-yl)urea

A reactor was charged with 1.00 mole-equivalents of N-methanesulfonylpiperid-4-yl amide, 1.06 mole-equivalents of 1-adamantyl amine, and 39.3mole-equivalents of acetonitrile, and the resulting mixture was heatedto 40° C. internal under a nitrogen atmosphere. (Diacetoxyiodo)benzene(1.20 mole-equivalents) was charged portionwise in such a way that thereaction mixture was maintained below 75° C. internal. After the(diacetoxyiodo)benzene had been added, the reaction mixture was heatedat 65-70° C. internal, and the reaction contents monitored until theamount of unreacted 1-adamantyl amine was less than 5% relative toproduct N-(1-methanesulfonyl piperidin-4-yl)-N′-(adamant-1-yl)urea(typically less than about 6 hours). The resulting mixture was cooled to20° C. internal and filtered to remove a small amount of insolublematerial. The filtrate was allowed to stand for 48 hours at which pointthe precipitated product was collected by filtration. The solid productwas dried to constant weight in a vacuum oven under a nitrogen bleedmaintaining an internal temperature of ≦50° C. to afford product in 58%yield based on N-methanesulfonyl piperid-4-yl amide. ¹H NMR(CDCl₃) δ:3.95-4.08 (m, 2H), 3.74-3.82 (m, 2H), 3.63-3.82 (m, 1H), 3.78 (s, 3H),3.70-3.80 (m, 2H), 2.02-2.12 (m, 5H), 1.90 (s, 6H), 1.67 (s, 6H),1.40-1.50 (m, 2H); MS: 356 [M+H]⁺; m.p. 228-229° C.

Example 43-63

Synthesized as described previously in Jones, P. D., et al. Bioorganic &medicinal chemistry letters 2006, 16, 5212.

Example 43

1-Piperidin-4-yl-3-(4-trifluoromethoxy-phenyl)-urea (1570)

¹H NMR (300 MHz, D6 DMS

) δ ppm 8.61 (s, 1H), 7.63-7.26 (m, 2H), 7.20 (d, J=8.29 Hz, 2H), 6.25(d, J=7.57 Hz, 1H), 3.60-3.38 (m, 1H), 2.87 (td, J=11.85, 3.18, 3.18 Hz,2H), 1.79-1.64 (m, 2H), 2.48-2.41 (m, 2H), 1.20 (qd, J=11.06, 3.83 Hz,2H); m.p. 169-173° C.

Example 44

1-(1-Acetyl-piperidin-4-yl)-3-(4-trifluoromethoxy-phenyl)-urea (1555)

¹H NMR (300 MHz, D6 DMS

) δ ppm 8.07 (s, 1H), 7.48-7.28 (m, 2H), 7.09 (d, J=8.93 Hz, 2H), 5.87(d, J=7.57 Hz, 1H), 4.53-4.26 (m, 1H), 4.05-3.82 (m, 1H), 3.82-3.70 (m,1H), 3.29-3.06 (m, 1H), 3.01-2.69 (m, 1H), 2.11 (s, 3H), 2.14-1.87 (m,2H), 1.40-1.25 (m, 2H); m.p. 198-202° C.

Example 45

1-[1-(2,2,2-Trifluoro-acetyl)-piperidin-4-yl]-3-(4-trifluoromethoxy-phenyl)-urea(1591)

¹H NMR (300 MHz, D6 DMS

) δ ppm 7.41-7.03 (m, 5H), 5.21 (s, 1H), 4.50-4.31 (m, 1H), 3.95 (brd,J=2H), 3.18 (m, 1H), 2.90 (m, 1H), 2.19-1.90 (m, 1H), 1.29 (m, 2H); m.p.150-154° C.

Example 46

1,3-Di-piperidin-4-yl-urea (1604)

¹H NMR (300 MHz, D6 DMS

) δ ppm 9.19-8.89 (m, 2H), 3.74-3.55 (m, 1H), 3.26-3.12 (m, 2H),2.95-2.80 (m, 2H), 1.99-1.79 (m, 2H), 1.65-1.45 (m, 2H).

Example 47

1,3-Bis-(1-benzoyl-piperidin-4-yl)-urea (1605)

¹H NMR (300 MHz, CDCl₃.) δ ppm 7.67-6.93 (m, 5), 5.39 (d, J=7.98 Hz,1H), 4.78-4.24 (m, 1H), 4.05-3.43 (m, 2H), 3.34-2.63 (m, 2H), 2.20-1.57(m, 2H), 1.49-0.71 (m, 2H); ¹³C NMR (75 MHz, CDCl₃)) δ ppm 170.84,157.25, 135.88, 130.14, 128.84, 126.94, 46.91, 46.66, 41.47, 33.97,32.67.

Example 48

Adamantane-1-carboxylic acid piperidin-4-ylamide

¹H NMR (300 MHz, CDCl₃.) δ ppm 8.97 (d, J=7.53 Hz, 2H), 7.39 (d, J=7.53Hz, 1H), 3.86-3.73 (m, 1H), 3.28-3.17 (m, 2H), 2.99-2.81 (m, 2H),2.01-1.53 (m, 19H).

Adamantane-1-carboxylic acid (1-acetyl-piperidin-4-yl)-amide (1641)

¹H NMR (300 MHz, CDCl₃.) δ ppm 5.49 (d, J=7.38 Hz, 1H), 4.60-4.51 (m,1H), 4.07-3.90 (m, 1H), 3.79 (ddd, J=13.76, 5.64, 3.68 Hz, 1H), 3.17(ddd, J=14.01, 12.08, 2.76 Hz, 1H), 2.79-2.67 (m, 1H), 2.10 (s, 3H),2.09-1.65 (m, 17H), 1.37-1.21 (m, 2H).

Example 49

2-Adamantan-1-yl-N-piperidin-4-yl-acetamide

¹H NMR (300 MHz, D6 DMS

) δ ppm 9.04-8.94 (m, 2H), 7.90 (d, J=7.49 Hz, 1H), 3.99-3.48 (m, 1H),3.24-3.12 (m, 2H), 3.00-2.79 (m, 2H), 2.10-1.22 (m, 21H).

N-(1-Acetyl-piperidin-4-yl)-2-adamantan-1-yl-acetamide (1642)

¹H NMR (300 MHz, CDCl₃) δ ppm 5.56 (d, J=7.73 Hz, 1H), 4.55 (d, J=14.17Hz, 1H), 4.08-3.93 (m, 1H), 3.84-3.73 (m, 1H), 3.22-3.10 (m, 1H),2.78-2.65 (m, 1H), 2.09 (s, 1H), 2.07-1.92 (m, 3H), 1.91 (s, 2H),1.76-1.55 (m, 12H), 1.39-1.21 (m, 2H).

Example 50

2-Adamantan-1-yl-N-[1-(2,2,2-trifluoro-acetyl)-piperidin-4-yl]-acetamide(1642)

¹H NMR (300 MHz, CDCl₃.) 8 ppm 5.30 (d, J=7.80 Hz, 1H), 4.56-4.46 (m,1H), 4.16-3.94 (m, 2H), 3.29-3.17 (m, 1H), 2.96-2.85 (m, 1H), 2.23-1.51(m, 19H), 1.47-1.31 (m, 2H).

Example 51

Adamantane-1-carboxylic acid[1-(2,2,2-trifluoro-acetyl)-piperidin-4-yl]-amide (1643)

¹H NMR (300 MHz, CDCl₃.) δ ppm 5.57-5.27 (m, 1H), 4.60-4.32 (m, 1H),4.18-3.85 (m, 2H), 3.36-3.09 (m, 1H), 3.00-2.77 (m, 1H), 2.17-1.52 (m,17H), 1.35 (s, 2H).

Example 52

1-(1-Acetyl-piperidin-4-yl)-3-cycloheptyl-urea (1645)

¹H NMR (300 MHz, CDCl₃.) δ ppm 4.68-4.60 (m, 2H), 4.49 (d, J=11.54 Hz,1H), 3.92-3.66 (m, 3H), 3.24-3.10 (m, 1H), 2.83-2.71 (m, 1H), 2.11 (s,3H), 2.08-1.11 (m, 16H).

Example 53

1-Adamantan-1-yl-3-(1-methanesulfonyl-piperidin-4-yl)-urea (1701)

¹H NMR (300 MHz, CDCl₃.) δ ppm 4.16-4.00 (m, 2H), 3.82-3.60 (m, 3H),2.78 (s, 3H), 2.76-2.69 (m, 2H), 2.12-1.61 (m, 17H), 1.53-1.36 (m, 2H).

Example 54

4-(3-Adamantan-1-yl-ureido)-piperidine-1-carboxylic acid methyl ester(1702)

¹H NMR (300 MHz, CDCl₃) δ ppm 5.68 (d, J=7.56 Hz, 1H), 5.42 (s, 1H),3.82-3.72 (m, 2H), 3.57 (s, 3H), 3.53-3.40 (m, 1H), 3.00-2.85 (m, 2H),2.03-1.52 (m, 17H), 1.20-1.04 (m, 2H).

Example 55

1-(1-Methanesulfonyl-piperidin-4-yl)-3-(4-trifluoromethoxy-phenyl)-urea(1709)

¹H NMR (300 MHz, CDCl₃.) δ ppm 8.04 (s, 1H), 7.44-7.38 (m, 2H), 7.09 (d,J=8.36 Hz, 2H), 5.93 (d, J=7.70 Hz, 1H), 4.00-3.55 (m, 3H), 2.90-2.78(m, 2H), 2.81 (s, 3H), 2.61-2.55 (m, 2H), 2.15-1.99 (m, 2H), 1.61-1.45(m, 2H).

Example 56

1-[1-(Toluene-4-sulfonyl)-piperidin-4-yl]-3-(4-trifluoromethoxy-phenyl)-urea(1711)

¹H NMR (300 MHz, CDCl₃.) δ ppm 7.91 (s, 1H), 7.63 (d, J=8.22 Hz, 2H),7.38-7.30 (m, 4H), 7.06 (d, J=8.84 Hz, 1H), 5.78 (s, 1H), 3.77-3.47 (m,3H), 2.48-2.35 (m, 2H), 2.44 (s, 3H), 2.05-1.93 (m, 2H), 1.57-1.41 (m,2H).

Example 57

1-[1-(5-Dimethylamino-naphthalene-1-sulfonyl)-piperidin-4-yl]-3-(4-trifluoromethoxy-phenyl)-urea(1710)

¹H NMR (300 MHz, CDCl₃.) δ ppm 8.59 (d, J=8.50 Hz, 1H), 8.32 (d, J=8.71Hz, 1H), 8.18 (dd, J=7.35, 1.19 Hz, 1H), 7.54 (ddd, J=8.50, 7.51, 3.20Hz, 2H), 7.29-7.22 (m, 2H), 7.19 (d, J=7.15 Hz, 1H), 7.02 (d, J=8.45 Hz,2H), 6.93 (s, 1H), 5.19 (d, J=7.73 Hz, 1H), 3.91-3.64 (m, 3H), 2.89 (s,6H), 2.86-2.72 (m, 2H), 2.06-1.92 (m, 2H), 1.64-1.46 (m, 2H).

Examples 58-64

These chemicals were synthesized by the direct reaction of amine withisocyanate following previously described procedures in Morisseau, C.,et al. Biochemical Pharmacology 2002, 63, 1599. Jones, P. D., et al.Bioorganic & medicinal chemistry letters 2006, 16, 5212.

Example 58

1-Cyclohexyl-3-(1,3,5-triaza-tricyclo[3.3.1.1^(3,7)]dec-7-yl)-urea(1549)

¹H NMR (300 MHz, D6 DMS

) δ ppm 5.62 (s, 1H), 5.37 (s, 1H), 4.25 (d, J=11.5 Hz, 3H), 3.90 (d,J=10.8 Hz, 3H), 3.50-3.21 (m, 6H), 2.50 (s, 1H), 1.82-1.59 (m, 5H)1.15-0.95 (m, 5H); m.p. 150-154° C.

Example 59

1-Dodecyl-3-piperidin-4-yl-urea (1550)

¹H NMR (300 MHz, D6 DMS

) δ ppm 8.79-8.35 (m, 3H), 3.74-3.55 (m, 1H), 3.26-3.12 (m, 4H),2.95-2.80 (m, 2H), 1.99-1.79 (m, 2H), 1.65-1.25 (m, 22H), 0.97 (t,J=12.8 Hz, 3H); m.p. 102-105° C.

Example 60

4-(3-Cyclohexyl-ureido)-piperidine-1-carboxylic acid tert-butyl ester(1551)

¹H NMR (300 MHz, CDCl₃) δ ppm 4.26-4.16 (t, J=8.6 Hz, 2H), 4.11-3.86 (m,2H), 3.80-3.64 (m, 1H), 3.55-3.39 (m, 1H), 2.94-2.78 (t, J=12.2 Hz, 2H),1.98-1.87 (m, 4H), 1.75-1.65 (m, 3H), 1.45-1.05 (m, 16H); m.p. 167-169°C.

Example 61

1-Dodecyl-3-(1H-indol-5-yl)-urea (1553)

¹H NMR (300 MHz, D6 DMS

) δ ppm 10.9 (s, 1H), 8.10 (s, 1H), 7.50 (s, 1H), 7.30-7.18 (m, 2H),7.00-6.90 (m, 1H), 6.19 (s, 1H), 6.00-5.95 (m, 1H), 3.41-3.18 (m, 2H),1.60-1.10 (m, 20H), 0.97 (t, J=12.8 Hz, 3H); m.p. 110-113° C.

Example 62

1-Cyclohexyl-3-(1H-indol-5-yl)-urea (1554)

¹H NMR (300 MHz, D6 DMS

) δ ppm 10.8 (s, 1H), 8.00 (s, 1H), 7.55 (s, 1H), 7.25-7.15 (m, 2H),6.95-6.87 (m, 1H), 6.15 (s, 1H), 5.95-5.90 (m, 1H), 2.58-2.42 (m, 1H),1.85-1.05 (m, 10H); m.p. 145-148° C.

Example 63

1-(1H-Benzoimidazol-5-yl)-3-dodecyl-urea (1568)

¹H NMR (300 MHz, D6 DMS

) δ ppm 8.35-8.25 (m, 2H), 7.35-7.20 (m, 2H), 6.65-6.50 (m, 1H), 5.404.85 (m, 2H), 3.38-3.21 (m, 2H), 1.40-1.1.15 (m, 20H), 0.85 (t, J=7.6Hz, 3H); m.p. 107-109° C.

Example 64

1-(1H-Benzoimidazol-5-yl)-3-cyclohexyl-urea (1569)

¹H NMR (300 MHz, D6 DMS

) δ ppm 8.41-8.29 (s, 1H), 8.15-8.00 (m, 1H), 7.35-7.15 (m, 2H),6.65-6.47 (m, 1H), 5.40-4.90 (m, 2H), 2.55-2.47 (m, 1H), 1.95-1.05 (m,10H); m.p. 143-148° C.

Example 65

Pyridin-4-ylmethyl-carbamic acid biphenyl-3-yl ester (1557)

White fine crystal. ¹H NMR (CDCl₃): 8.60 (d, J=5.70 Hz, 2H), 7.10-7.50(m, 11H), 5.56 (br, 1H), 4.50 (d, J=6.30 Hz, 2H); m.p.: 132° C.

Example 66

Pyridin-4-ylmethyl-carbamic acid 2-methyl-biphenyl-3-ylmethyl ester(1558)

White sponge-like crystal, ¹H NMR (CDCl₃): 8.56 (d, J=5.70 Hz, 2H),7.20-7.45 (m, 10H), 5.25 (s, 3H), 4.42 (d, J=6.30 Hz, 2H), 2.25 (s, 3H);m.p. 103° C.

Example 67

Pyridin-3-ylmethyl-carbamic acid biphenyl-3-yl ester (1559)

White crystal, ¹H NMR (CDCl₃): 8.62 (s, 1H), 8.57 (dd, J₁=1.20 Hz,J₂=4.50 Hz, 1H), 7.70-7.75 (m, 11H), 7.56-7.59 (m, 12H), 7.27-7.46 (m,7H), 7.11-7.15(m, 1H), 5.54 (br, 1H), 4.49 (d, J=6.30 Hz, 2H); m.p. 113°C.

Example 68

Pyridin-3-ylmethyl-carbamic acid 2-methyl-biphenyl-3-ylmethyl ester(1560)

White crystal, ¹H NMR (CDCl₃): 8.55 (m, 2H), 7.76 (d, J=7.50 Hz, 1H),7.22-7.41 (m, 9H), 5.23 (s, 2H), 5.19 (br, 1H), 4.42 (d, J=5.70 Hz, 2H),2.23 (s, 3H); m.p. 110° C.

Example 69

Morpholine-4-carboxylic acid biphenyl-3-yl ester (1561)

White crystal, ¹H NMR (CDCl₃): 7.57-7.60 (m, 2H), 7.27-7.46 (m, 6H),7.08-7.13 (m, 1H), 3.60-3.79 (m, 8H); m.p. 97° C.

Example 70

Morpholine-4-carboxylic acid 2-methyl-biphenyl-3-ylmethyl ester (1562)

Colorless sticky oil. ¹H NMR (CDCl₃): 7.23-7.45 (m, 8H), 5.23 (s, 2H),3.68 (br, 4H), 3.50-3.53(m, 4H), 2.23 (s, 3H).

Example 71

This example provides assays and illustrates the inhibition of humansoluble epoxide hydrolases by compounds of the invention.

Enzyme Preparation

Recombinant human sEH was produced in a baculovirus expression systemand purified by affinity chromatography. The preparations were at least97% pure as judged by SDS-PAGE and scanning densitometry. No detectableesterase or glutathione transferase activity, which can interfere withthis sEH assay, was observed. Protein concentration was quantified byusing the Pierce BCA assay using Fraction V bovine serum albumin as thecalibrating standard.

IC₅₀ Assay Conditions

IC₅₀ values were determined in one of three methods. One method usesracemic 4-nitrophenyl-trans-2,3-epoxy-3-phenylpropyl carbonate assubstrate. Enzyme (0.24 μM human sEH) was incubated with inhibitors for5 min in sodium phosphate buffer, 0.1 M pH 7.4, at 30° C. beforesubstrate introduction ([S]=40 μM). Activity was assessed by measuringthe appearance of the 4-nitrophenolate anion at 405 nm at 30° C. during1 min (Spectramax 200; Molecular Devices). Assays were performed intriplicate. IC₅₀ is a concentration of inhibitor, which reduces enzymeactivity by 50%, and was determined by regression of at least five datumpoints with a minimum of two points in the linear region of the curve oneither side of the IC₅₀. The curve was generated from at least threeseparate runs, each in triplicate.

Other IC₅₀ values were determined using the procedure described inAnalytical Biochemistry 343 66-75 (2005) usingcyano(6-methoxy-naphthalen-2-yl)methyltrans-[(3-phenyloxiran-2-yl)methyl]carbonate as a substrate. Enzymes(0.96 nM for human sEH) were incubated with inhibitors ([I]=0.5-10,000nM) for 5 min in BisTris-HCl buffer (25 mM, pH 7.0, containing 0.1 mg/mlof BSA) at 30° C. prior to substrate introduction ([S]=51M). Enzymeactivity was measured by monitoring the appearance of6-methoxy-2-naphthaldehyde. Assays were performed in triplicate. Bydefinition, IC₅₀ values are concentrations of inhibitor that reduceenzyme activity by 50%. IC₅₀ values were determined by regression of atleast five datum points, with a minimum of two datum points in thelinear region of the curve on either side of the IC₅₀ values. The curvewas generated from at least three separate runs, each in triplicate.

Other inhibition potencies were determined using a fluorescent basedhigh-throughput assay. Inhibitors in solution at 10 mM in DMSO wereserially diluted by 10-fold increment in Bis/Tris HCl buffer (25 mM pH7.0) containing 0.1 mg/mL of BSA (Buffer A). In black 96-well plates, 20μL of the inhibitor dilution or buffer were delivered in every well, andthen 130 μL of Human sEH at ˜0.4 μg/mL in solution in Buffer A wereadded to each well. The plate was then mixed and incubated at roomtemperature for 5 minutes. Fifty microliters of substrate((3-Phenyl-oxiranyl)-acetic acidcyano-(6-methoxy-naphthalen-2-yl)-methyl ester; PHOME) at 200 μM insolution in 96:4 Buffer A:DMSO was then added to each well to give[S]final=50 μM and [E]final=˜4 nM. The plate was then mixed andincubated in the dark at room temperature (˜25° C.) for 90 min. Activitywas measured by determining the relative quantity of6-methoxy-2-naphthaldehyde formed with an excitation wavelength of 316nm and an emission wavelength of 460 nm measured with a SpectraMax M-2fluorometer (molecular Devices, Sunnyvale Calif.).

Assays were conducted with the compounds indicated in Table 1-5, asdescribed above.

Example 72

R Compound # IC₅₀ (nM)

H 1175 960 Et 1152 3758 n-Pr 1155 809 n-Bu 1160 1249 benzyl 1158 8.4

H 1118 4258 Et 1154 3949 n-Pr 1122 2578 n-Bu 1161 613 benzyl 1119 112

This example illustrates the inhibition of human soluble epoxidehydrolases by compounds of the invention having an alkyl substitutedpiperidine moiety.

Assays were conducted with the compounds indicated in Table 1, accordingto established protocols (see, above).

Table 1: Inhibition of Human sEH by Alkyl Substituted Piperidines:

Example 73

This example illustrates the inhibition of human soluble epoxidehydrolases by compounds of the invention having an amide substitutedpiperidine moiety.

Assays were conducted with the compounds indicated in Table 2, accordingto established protocols (see, above).

TABLE 2 Inhibition of human sEH by simple amide substituted piperidines:R Compound # IC₅₀ (nM)

—C(O)Me 1153 14.5 —C(O)Et 1163 3.2 —C(O)-n-Pr 1157 2.6 —C(O)Ph 1159 1.3

1201 1.2

1433 1.7

1434 2.1

—C(O)Me 1156 5.0 —C(O)Et 1162 8.7 —C(O)-n-Pr 1120 6.7 —C(O)Ph 1121 3.2

1207 7.6

1435 5.4

1436 7.3

Example 74

This example illustrates the inhibition of human soluble epoxidehydrolases by compounds of the invention having an amide-estersubstituted piperidine moiety.

Assays were conducted with the compounds indicated in Table 3, accordingto established protocols (see, above).

TABLE 3 Inhibition of human sEH by amide-ester piperidines: R Compound #IC₅₀ (nM)

1205 9.0

1206 2.7

1202 1.7

1203 1.1

1204 1.1

1208 6.2

1212 3.4

1210 1.8

1209 4.1

1211 1.5

Example 75

This example illustrates the inhibition of human soluble epoxidehydrolases by compounds of the invention having an amide-acidsubstituted piperidine moiety.

Assays were conducted with the compounds indicated in Table 4, accordingto established protocols (see, above).

TABLE 4 Inhibition of human sEH by amide-acid piperidines: R Compound #IC₅₀ (nM)

1503 254.5

1501 72.8

1507 161.2

1505 10.1

1523 3.3

1502 174.5

1500 41.6

1506 407.1

1504 43.6

1522 11.8

Example 76

This example provides a table of structures of compounds with variousother functionalities included in the invention. For example, the ureapharmacophore can be varied with amide or carbamate functionality toimprove physical properties of sEH inhibitors as shown in Table 5a.

Assays were conducted with the compounds indicated in Table 5a and 5b,according to established protocols (see, above).

TABLE 5a Inhibition of human sEH by1-substituted-3-n-(substituted)heterocyclic ureas, carbamates andamides: Compound Structure IC₅₀ (nM) 1549

13586.4 1550

42.5 1551

4.3 1553

639.1 1554

87 1555

11.5 1556

1.8 1557

11468.1 1558

1329.7 1559

22991.5 1560

4413.4 1561

65339.4 1562

11994.1 1567

1568

5021 1569

457 1570

2316 1590

1.1 1591

0.4 1602

561.7 1604

100000 1605

4.8 1606

1.7 1641

12649.3 1642

275.1 1643

4208.9 1644

28.3 1645

27.6

TABLE 5b Structure Compound # IC50 (nM)

1701 1.4

1702 0.9

1710 0.8

1711 0.4

Example 77

Pharmacokinetic Screening Procedure:

This example provides the pharmacokinetic studies, specifically serumprofiles carried out using sEH inhibitorory compounds of the presentinvention in dogs. As noted above, the use of 1-substituted ureainhibitors afforded exquisite sensitivity, allowing the determination ofthe determined pharmacokinetic parameters from serial blood samplescollected from individual dogs (see Tables 6-8).

Animals.

Healthy dogs, 5-6 years-old, were assigned to study groups based onbody-weight stratified randomization procedure. The body weight ofanimals used in all the experiments was about 20 kg. Dogs weremaintained on a natural light/dark cycle under standard kennelconditions, with food and water available ad libid um.

Drug Preparation, Administration, Blood Sample Drawing.

Various amounts of a compound was dissolved in 1 mL of Crisco, heatedand sonicated for 15 minutes to dissolve the compounds. The mixture wastransferred in solution to a syringe with a cap. The mixture becomes asolid at room temperature and may be kept in a refrigerator until used.sEH inhibitors were administered orally to dogs via syringe. Thecompounds are administered at room temperature or warmer so that theyare in preferably in solution. The dogs are fed immediately thereafter.

Serial blood samples (100 μL) were collected from a catheter inserted inthe right front leg of the dog. Serial blood samples were collected inEDTA tubes at various time points (0, 15, 30, 60, 120, 180, 240, 300,360, 480, and 1440 minutes) after administration. The blood samples arecentrifuged at 4000 rpm for 10 minutes and the plasma is collected intomicro-centrifuge tubes and frozen at −80° C.

Plasma Sample Preparation for LC/MS Measurement and Analysis

100 uL of plasma was collected in another Eppendorf. 200 μL of water and500 μL of ethyl acetate is added and the mixture was vortexed. 10 uL ofsurrogate was added and the mixture was vortexed again. The mixture wascentrifuged for 6000 rpm for 5 min. and the organic phase was thenextracted into another Eppendorf. Another 500 uL of ethyl acetate isadded to the water phase and the mixture is extracted again. The organicphase is dried under nitrogen and the samples reconstituted with 50 μLof MeOH and at least one internal standard is added to the plasmamixture (e.g. an extraction standard). Aliquots (5 μL) were injectedonto the LC-MS/MS system. For measuring parent compounds and theirmetabolites by using LC-MS/MS: a Waters 2790 liquid chromatographequipped with a 30×2.1 mm 3 μm C18 Xterra™ column (Waters) and aMicromass Quattro Ultima triple quadrupole tandem mass spectrometer(Micromass, Manchester, UK) was used.

Analysis.

Pharmacokinetics analysis was performed using SigmaPlot software system(SPSS science, Chicago, Ill.). A one-compartment model was used forblood concentration-time profiles for the oral gavage dosing and fits tothe following equation (see, Gibson, G. G. and Skett, P.: INTRODUCTIONTO DRUG METABOLISM, SECOND ED., Chapman and Hall, New York 1994,199-210):C=ae ^(−bt)The half-life (t_(1/2)) for the elimination phase was calculated by thefollowing equation:t _(1/2)=0.693/bThe area under the concentration (AUC) was calculated by the followingequation:AUC=a/bWhere:

-   -   C=the total blood concentration at time t    -   a=the extrapolated zero intercept    -   b=the apparent first-order elimination rate constant

The results shown in Tables 6, 7 and 8 and examples of the time courseof compounds is shown in FIGS. 1-3.

TABLE 6 LC/MS analysis Compound Time 1153 1555 1606 1163 1157 1159 11211201 1204 1206 0 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 0.00 15288.97 16.07 0.00 5.70 0.58 7.26 4.86 9.19 0.00 0.00 30 276.16 45.761.61 16.05 5.18 7.82 3.81 32.65 0.19 0.00 60 241.03 100.35 4.82 31.797.77 4.58 1.98 42.48 0.34 0.00 120 107.19 216.91 9.37 24.00 9.50 1.390.42 21.09 0.11 0.00 180 56.41 260.79 10.98 14.55 7.05 0.60 0.16 7.150.00 0.00 240 26.42 268.90 6.83 10.05 4.60 0.00 0.06 3.16 0.00 0.00 30020.81 302.64 5.62 7.20 2.45 0.00 0.05 1.72 0.00 0.00 360 10.10 3.21 4.952.45 0.00 0.03 0.00 0.00 0.00 480 4.66 320.45 1.47 1.50 0.86 0.00 0.030.00 0.00 0.00 1440 0.00 266.29 0.00 0.00 0.00 0.04 0.00 0.00 0.00

Note the acid moiety makes a big difference in these compounds. Theacids reach a maximum concentration faster and gives sustained bloodlevels. Higher blood levels (bioavailability) generally correspond tohigher protein binding and higher efficacy. Accordingly, the presence ofan acidic moiety improves the oral availability of these inhibitors.

TABLE 7 Pharmacokinetic parameters of compounds AUCINF_D_(—)AUCINF_D_(—) Cmpd. AUC pred pred/ (solubility IC₅₀ (μM* (min*kg* IC₅₀ inoil) Structure (nM) min) nM/mg) (AUC/IC₅₀) 1153 (Opaque)

14.5 35.8 119214 8221 (2.5) 1156

5 4.9 (1.0) 1555 (Opaque)

11.5 390.1 13002825 1130680 (33.9) 1606 (Suspension)

1.7 2.6 8553 5031 (1.5) 1163 (Opaque)

3.19 5.6 18504 5800 (1.75 1157 (Opaque)

2.64 2.2 7187 2722 (0.8) 1159 (Opaque)

1.3 1121 (Not dissolved)

3.2 1201 (Suspension)

1.2 4.8 16100 13416 (4.0) 1204 (Opaque)

1.1 1206 (Opaque)

2.7 1642

275.1 5.8 (0.02) 1644

28.3 1645

27.6 220.7 (8.0) 1701

1.4 4.9 (3.5) 1702

0.9 5.4 (6.0) 1710

0.8 1711

0.4

TABLE 8 Compound 1153 in Compound 1153 in Time solution at 0.1 mg/kgdose solution at 0.3 mg/kg dose 0 0.31 0.00 15 5.33 288.97 30 19.59276.16 60 56.74 241.03 120 106.43 107.19 180 100.94 56.41 240 75.7126.42 300 44.51 20.81 360 29.15 10.10 480 13.01 4.66 1440 0.00 0.00

1. A compound having the formula:

wherein R¹ is a member selected from the group consisting of phenyl,cyclohexyl, cycloheptyl and adamantyl, wherein said phenyl is optionallysubstituted with from 1 to 2 substituents each independently selectedfrom the group consisting of C₁-C₈alkyl, haloC₁-C₈alkyl, cyano,haloC₁-C₈alkoxy, C₁-C₈heteroalkyl, aryl, heteroaryl; L is selected fromthe group consisting of —CO— and —SO₂-; R⁴ is selected from the groupconsisting of H, C₁-C₈alkyl, arylC₀-C₈alkyl, and C₃-C₁₂cycloalkyl, eachoptionally substituted with from 1 to 2 substituents each independentlyselected from the group consisting of C₁-C₈alkyl, halo,C₁-C₈heteroalkyl, arylC₀-C₈alkyl, COR⁶, S(O)_(m)R⁶ and heteroaryl; eachR⁶ is independently selected from the group consisting of H, C₁-C₈alkyl,C₁-C₈alkoxy and amino; the subscript n is 0; and the subscript m is aninteger of from 0 to 2; or a pharmaceutically acceptable salt thereof.2. The compound in accordance with claim 1 having the formula:

or a pharmaceutically acceptable salt thereof.
 3. The compound inaccordance with claim 1 or 2, wherein R¹ is adamantyl.
 4. The compoundin accordance with claim 1 or 2, wherein R¹ is cycloheptyl orcyclohexyl.
 5. The compound in accordance with claim 1 or 2, wherein Lis a —C(O)—.
 6. The compound in accordance with claim 1 or 2, wherein R⁴is selected from the group consisting of hydrogen, C₁-C₈ alkyl, andarylC₀-C₈alkyl.
 7. The compound in accordance with claim 1 or 2, whereinR⁴ is C₁-C₈ alkyl.
 8. A compound according to claim 2 which compound isselected from the group consisting of:N-(1-acetylpiperidin-4-yl)-N′-(adamant-1-yl)urea;N-(1-propionylpiperidin-4-yl)-N′-(adamant-1-yl)urea;N-(1-butyrylpiperidin-4-yl)-N′-(adamant-1-yl)urea;N-(1-benzoylpiperidin-4-yl)-N′-(adamant-1-yl)urea;4-[4-(3-adamantan-1-yl-ureido)-piperidin-1-yl]-4-oxo-butanoic acidmethyl ester;5-[4-(3-adamantan-1-yl-ureido)-piperidin-1-yl]-5-oxo-pentanoic acidmethyl ester;2-[4-(3-adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acidmethyl ester;3-[4-(3-adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acidmethyl ester;4-[4-(3-adamantan-1-yl-ureido)-piperidine-1-carbonyl]-benzoic acidmethyl ester;1-(1-acetyl-piperidin-4-yl)-3-(4-trifluoromethoxyphenyl)urea;1-(1-trifluoromethylcarbonylpiperidin-4-yl)-3-(4-trifluoromethoxyphenyl)urea;and 1-(1-acetyl-piperidin-4-yl)-3-cycloheptyl urea; or apharmaceutically acceptable salt thereof.
 9. The compound according toclaim 8, wherein said compound is selected from the group consisting of:N-(1-acetylpiperidin-4-yl)-N′-(adamant-1-yl)urea;N-(1-propionylpiperidin-4-yl)-N′-(adamant-1-yl)urea;N-(1-butyrylpiperidin-4-yl)-N′-(adamant-1-yl)urea; andN-(1-benzoylpiperidin-4-yl)-N′-(adamant-1-yl)urea or a pharmaceuticallyacceptable salt thereof.
 10. The compound according to claim 8, whereinsaid compound is N-(1-acetylpiperidin-4-yl)-N′-(adamant-1-yl)urea or apharmaceutically acceptable salt thereof.
 11. The compound according toclaim 8, wherein said compound isN-(1-propionylpiperidin-4-yl)-N′-(adamant-1-yl)urea or apharmaceutically acceptable salt thereof.
 12. The compound according toclaim 8, wherein said compound isN-(1-butyrylpiperidin-4-yl)-N′-(adamant-1-yl)urea or a pharmaceuticallyacceptable salt thereof.
 13. The compound according to claim 8, whereinsaid compound is N-(1-benzoylpiperidin-4-yl)-N′-(adamant-1-yl)urea or apharmaceutically acceptable salt thereof.
 14. A compound according toclaim 2 which compound is selected from the group consisting of:N-(1-methanesulfonylpiperidin-4-yl)-N′-(adamant-1-yl)urea;N-(1-methanesulfonylpiperidin-4-yl)-N′-(4-trifluoromethoxyphenyl)urea;N-(1-toluene-4-sulfonylpiperidin-4-yl)-N′-(4-trifluoromethoxyphenyl)urea;andN-[1-(5-dimethylaminonaphthalene-1-sulfonyl)-piperidin-4-yl]-N′-(4-trifluoromethoxy-phenyl) urea; or a pharmaceutically acceptable salt thereof.
 15. Thecompound according to claim 14, wherein said compound isN-(1-methanesulfonylpiperidin-4-yl)-N′-(adamant-1-yl)urea or apharmaceutically acceptable salt thereof.
 16. The compound according toclaim 14, wherein said compound isN-(1-methanesulfonylpiperidin-4-yl)-N′-(4-trifluoromethoxy-phenyl)ureaor a pharmaceutically acceptable salt thereof.
 17. The compoundaccording to claim 14, wherein said compound isN-(1-toluene-4-sulfonylpiperidin-4-yl)-N′-(4-trifluoromethoxy-phenyl)ureaor a pharmaceutically acceptable salt thereof.
 18. The compoundaccording to claim 14, wherein said compound isN-[1-(5-dimethylaminonaphthalene-1-sulfonyl)-piperidin-4-yl]-N′-(4-trifluoromethoxy-phenyl)urea or a pharmaceutically acceptable salt thereof.
 19. A pharmaceuticalcomposition comprising a compound according to any one of claims 1 and 2and a pharmaceutically acceptable excipient.